U.S. patent application number 17/625599 was filed with the patent office on 2022-08-25 for oxazole compound as multi-targeted inhibitor of irak4 and btk.
The applicant listed for this patent is MEDSHINE DISCOVERY INC.. Invention is credited to Shuhui CHEN, Jian LI, Jie LI, Haizhong TAN, Jianfei WANG, Yang ZHANG.
Application Number | 20220267322 17/625599 |
Document ID | / |
Family ID | |
Filed Date | 2022-08-25 |
United States Patent
Application |
20220267322 |
Kind Code |
A1 |
WANG; Jianfei ; et
al. |
August 25, 2022 |
OXAZOLE COMPOUND AS MULTI-TARGETED INHIBITOR OF IRAK4 AND BTK
Abstract
Provided are a class of multi-targeted inhibitors of IRAK4 and
BTK, and the use thereof in preparing a drug for treating IRAK4-
and BTK-related diseases. The present invention specifically
relates to the compounds represented by formula (II), isomers
thereof or pharmaceutically acceptable salts thereof.
##STR00001##
Inventors: |
WANG; Jianfei; (Shanghai,
CN) ; TAN; Haizhong; (Shanghai, CN) ; LI;
Jie; (Shanghai, CN) ; ZHANG; Yang; (Shanghai,
CN) ; LI; Jian; (Shanghai, CN) ; CHEN;
Shuhui; (Shanghai, CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
MEDSHINE DISCOVERY INC. |
|
|
|
|
|
Appl. No.: |
17/625599 |
Filed: |
July 10, 2020 |
PCT Filed: |
July 10, 2020 |
PCT NO: |
PCT/CN2020/101369 |
371 Date: |
January 7, 2022 |
International
Class: |
C07D 471/04 20060101
C07D471/04; A61P 35/00 20060101 A61P035/00 |
Foreign Application Data
Date |
Code |
Application Number |
Jul 10, 2019 |
CN |
201910619602.8 |
Dec 6, 2019 |
CN |
201911240843.8 |
May 28, 2020 |
CN |
202010470469.7 |
Claims
1. A compound represented by formula (II), an isomer thereof or a
pharmaceutically acceptable salt thereof, ##STR00147## wherein,
R.sub.1 is selected from H, F, Cl, Br, I, OH, NH.sub.2, CN,
C.sub.1-6 alkyl, cyclopropyl and --C(.dbd.O)--NH.sub.2, wherein the
C.sub.1-6 alkyl, cyclopropyl and --C(.dbd.O)--NH.sub.2 are
optionally substituted by 1, 2 or 3 R.sub.a; R.sub.2 is selected
from thienyl, phenyl, pyridyl, cyclopropyl, cyclohexyl and
##STR00148## wherein the thienyl, phenyl, pyridyl, cyclopropyl,
cyclohexyl and ##STR00149## are optionally substituted by 1, 2, 3,
4 or 5 R.sub.b; T.sub.1 is selected from CH.sub.2, NH and O;
R.sub.3 is selected from C.sub.1-6 alkyl, wherein the C.sub.1-6
alkyl is optionally substituted by 1, 2 or 3 R.sub.c; R.sub.a is
each independently selected from F, OH, NH.sub.2 and CN; R.sub.b is
each independently selected from H, D, F, Cl, Br, I, OH, NH.sub.2,
CN, C.sub.1-3 alkyl, COOH, --C(.dbd.O)--C.sub.1-3 alkyl,
--C(.dbd.O)--O--C.sub.1-3 alkyl and --C(.dbd.O)--NH.sub.2, wherein
the OH, NH.sub.2, C.sub.1-3 alkyl, COOH, --C(.dbd.O)--C.sub.1-3
alkyl, --C(.dbd.O)--O--C.sub.1-3 alkyl and --C(.dbd.O)--NH.sub.2
are optionally substituted by 1, 2 or 3 R; R.sub.c is each
independently selected from F, OH, NH.sub.2, CN, CH.sub.3, COOH and
--SO.sub.2CH.sub.3; R is each independently selected from F, OH,
NH.sub.2 and CH.sub.3.
2. The compound as defined in claim 1, the isomer thereof or the
pharmaceutically acceptable salt thereof, wherein, R.sub.1 is
selected from H, F, Cl, Br, I, OH, NH.sub.2, CN, C.sub.1-3 alkyl,
cyclopropyl and --C(.dbd.O)--NH.sub.2, wherein the C.sub.1-3 alkyl,
cyclopropyl and --C(.dbd.O)--NH.sub.2 are optionally substituted by
1, 2 or 3 R.sub.a.
3. The compound as defined in claim 2, the isomer thereof or the
pharmaceutically acceptable salt thereof, wherein, R.sub.1 is
selected from CN, CH.sub.3, CF.sub.3, ##STR00150## and
--C(.dbd.O)--NH.sub.2.
4. The compound as defined in claim 1, the isomer thereof or the
pharmaceutically acceptable salt thereof, wherein, R.sub.b is each
independently selected from H, D, F, Cl, Br, I, OH, NH.sub.2, CN,
CH.sub.3, CH.sub.2CH.sub.3, CH.sub.2CH.sub.2CH.sub.3,
CH(CH.sub.3).sub.2, COOH, ##STR00151## wherein the OH, NH.sub.2,
CH.sub.3, CH.sub.2CH.sub.3, CH.sub.2CH.sub.2CH.sub.3,
CH(CH.sub.3).sub.2, ##STR00152## are optionally substituted by 1, 2
or 3 R.
5. The compound as defined in claim 4, the isomer thereof or the
pharmaceutically acceptable salt thereof, wherein, R.sub.b is each
independently selected from H, D, F, Cl, OH, OCH.sub.3, CN,
CH.sub.3, CH.sub.2OH, CH.sub.2NH.sub.2, COOH, ##STR00153##
6. The compound as defined in claim 1, the isomer thereof or the
pharmaceutically acceptable salt thereof, wherein, R.sub.2 is
selected from thienyl, phenyl, pyridyl, cyclopropyl, cyclohexyl,
##STR00154## wherein the thienyl, phenyl, pyridyl, cyclopropyl,
cyclohexyl, ##STR00155## are optionally substituted by 1, 2, 3, 4
or 5 R.sub.b.
7. The compound as defined in claim 6, the isomer thereof or the
pharmaceutically acceptable salt thereof, wherein, R.sub.2 is
selected from ##STR00156##
8. The compound as defined in claim 1, the isomer thereof or the
pharmaceutically acceptable salt thereof, wherein, R.sub.2 is
selected from ##STR00157##
9. The compound as defined in claim 1, the isomer thereof or the
pharmaceutically acceptable salt thereof, wherein, R.sub.3 is
selected from C.sub.2-5 alkyl, wherein the C.sub.2-5 alkyl is
optionally substituted by 1, 2 or 3 R.sub.c.
10. The compound as defined in claim 9, the isomer thereof or the
pharmaceutically acceptable salt thereof, wherein, R.sub.3 is
selected from CH.sub.2CH.sub.3, CH.sub.2CH.sub.2CH.sub.3,
CH(CH.sub.3).sub.2, CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
CH.sub.2CH(CH.sub.3).sub.2 and CH.sub.2CH.sub.2CH(CH.sub.3).sub.2,
wherein the CH.sub.2CH.sub.3, CH.sub.2CH.sub.2CH.sub.3,
CH(CH.sub.3).sub.2, CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
CH.sub.2CH(CH.sub.3).sub.2 and CH.sub.2CH.sub.2CH(CH.sub.3).sub.2
are optionally substituted by 1, 2 or 3 R.sub.c.
11. The compound as defined in claim 10, the isomer thereof or the
pharmaceutically acceptable salt thereof, wherein, R.sub.3 is
selected from ##STR00158##
12. The compound as defined in claim 1, the isomer thereof or the
pharmaceutically acceptable salt thereof, the compound is selected
from, ##STR00159## ##STR00160## wherein, L.sub.1 is selected from
C.sub.2-5 alkyl, and R.sub.1, R.sub.3 and R.sub.b are as defined in
claim 1.
13. A compound represented by the following formula, an isomer
thereof or a pharmaceutically acceptable salt thereof, ##STR00161##
##STR00162## ##STR00163## ##STR00164## ##STR00165## ##STR00166##
##STR00167## ##STR00168##
14. A pharmaceutical composition, comprising a therapeutically
effective amount of the compound as defined in claim 1, the isomer
thereof or the pharmaceutically acceptable salt thereof as active
ingredient and a pharmaceutically acceptable carrier.
15. A method for treating diseases related to IRAK4 and BTK in a
subject in need thereof, comprising administering an effective
amount o the compound as defined in claim 1, the isomer thereof or
the pharmaceutically acceptable salt thereof to the subject.
16. A method for treating diseases related to IRAK4 and BTK in a
subject in need thereof, comprising administering an effective
amount of the pharmaceutical composition as defined in claim 14 to
the subject.
17. A method for inhibiting IRAK4 and/or BTK in a subject in need
thereof, comprising administering an effective amount of the
compound as defined in claim 1, the isomer thereof or the
pharmaceutically acceptable salt thereof to the subject.
18. A method for inhibiting IRAK4 and/or BTK in a subject in need
thereof, comprising administering an effective amount of the
pharmaceutical composition as defined in claim 14 to the subject to
the subject.
Description
THE PRESENT APPLICATION CLAIMS THE FOLLOWING PRIORITIES
[0001] CN201910619602.8, filed on Jul. 10, 2019; [0002]
CN201911240843.8, filed on Dec. 6, 2019; [0003] CN202010470469.7,
filed on May 28, 2020.
TECHNICAL FIELD
[0004] The present disclosure relates to a class of multi-targeted
inhibitors of IRAK4 and BTK, and a use thereof in the preparation
of a medicament for treating IRAK4- and BTK-related diseases. The
present disclosure specifically relates to a compound represented
by formula (II), an isomer thereof or a pharmaceutically acceptable
salt thereof.
PRIOR ART
[0005] Interleukin-1 receptor-associated kinase 4 (IRAK4) is a
serine/threonine-specific protein kinase, a member of the tyrosine
like kinase (TLK) family, and a key node in the innate immune
response involving interleukin-1, 18, 33 receptors and Toll-like
receptors. After binding with interleukin receptor or Toll-like
receptor, extracellular signal molecules recruit to form MyD88:
IRAK4: IRAK1/2 multiprotein complex, leading to phosphorylation of
IRAK1/2 and mediating a series of downstream signal transduction,
thus activating p38, JNK and NF-.kappa. B signaling pathways, and
finally leading to the expression of proinflammatory cytokines.
Clinicopathological studies have shown that individuals with IRAK4
mutation have a protective effect on chronic lung disease and
inflammatory bowel disease. IRAK4 deficiency itself is non-lethal,
individuals can survive to adulthood, and the risk of infection
decreases with age. Therefore, IRAK4 has become an important
therapeutic target, which can be widely used in the treatment of
inflammatory diseases, immune diseases, tumor diseases and other
diseases. As shown in the following figure, BAY-1830839 and
BAY-1834845 are small molecule IRAK4 inhibitors developed by Bayer
Company, at present, clinical research on immune and tumor diseases
has been carried out.
##STR00002##
[0006] Activated B-cell-like diffuse large B-cell lymphoma
(ABC-DLBCL) is a highly invasive and poorly prognostic DLBCL, which
is usually characterized by abnormalities of B-cell receptor (BCR)
pathway and myeloid-like differentiation factor 88 (MyD88) pathway,
which further leads to the continuous activation of nuclear factor
.kappa.B protein (NF-.kappa.B) signaling pathway. CD79 mutation is
a common abnormal mutation in BCR pathway, and BTK inhibitors such
as Ibrutinib can inhibit the abnormal activation of NF-.kappa. B
signaling pathway caused by CD79 mutation, thus inhibiting the
proliferation of ABC-DLBCL cells. The abnormal MyD88 pathway is
mainly MyD88.sup.L265P point mutation, which accounts for about
30%, IRAK4 inhibitors can effectively block the abnormally
activated MyD88 signaling pathway and further block the abnormal
activation of the NF-.kappa.B pathway. However, ABC-DLBCL patients
with MyD88.sup.L265P mutations have a poor response to BCR
inhibitors due to abnormal MyD88 signaling pathway, and a large
number of research data from Bayer, Nimbus and AstraZeneca indicate
that the combination of IRAK4 inhibitor and BTK inhibitor can
significantly improve the in vivo efficacy of Ibrutinib in
ABC-DLBCL xenotransplantation animal model. If the abnormality of
BCR pathway and MyD88 pathway can be effectively inhibited at the
same time, it will be a more effective way to treat ABC-DLBCL,
therefore, developing RAK4 and BTK dual-target inhibitors can
obtain double benefits in blocking NF-.kappa.(B pathway, which is a
very efficient and effective strategy in terms of therapeutic
mechanism and provides a potentially effective new therapeutic
method for ABC-DLBCL patients.
CONTENT OF THE PRESENT INVENTION
[0007] The present disclosure provides a compound represented by
formula (II), an isomer thereof or a pharmaceutically acceptable
salt thereof,
##STR00003##
[0008] wherein,
[0009] R.sub.1 is selected from H, F, Cl, Br, I, OH, NH.sub.2, CN,
C.sub.1-6 alkyl, cyclopropyl and --C(.dbd.O)--NH.sub.2, wherein the
C.sub.1-6 alkyl, cyclopropyl and --C(.dbd.O)--NH.sub.2 are
optionally substituted by 1, 2 or 3 R.sub.a;
[0010] R.sub.2 is selected from thienyl, phenyl, pyridyl,
cyclopropyl, cyclohexyl and
##STR00004##
wherein the thienyl, phenyl, pyridyl, cyclopropyl, cyclohexyl
and
##STR00005##
are optionally substituted by 1, 2, 3, 4 or 5 R.sub.b;
[0011] T.sub.1 is selected from CH.sub.2, NH and O;
[0012] R.sub.3 is selected from C.sub.1-6 alkyl, wherein the
C.sub.1-6 alkyl is optionally substituted by 1, 2 or 3 R.sub.c;
[0013] R.sub.a is each independently selected from F, OH, NH.sub.2
and CN;
[0014] R.sub.b is each independently selected from H, D, F, Cl, Br,
I, OH, NH.sub.2, CN, C.sub.1-3 alkyl, COOH, --C(.dbd.O)--C.sub.1-3
alkyl, --C(.dbd.O)--O--C.sub.1-3 alkyl and --C(.dbd.O)--NH.sub.2,
wherein the OH, NH.sub.2, C.sub.1-3 alkyl, --C(.dbd.O)--C.sub.1-3
alkyl, --C(.dbd.O)--O--C.sub.1-3 alkyl and --C(.dbd.O)--NH.sub.2
are optionally substituted by 1, 2 or 3 R;
[0015] R.sub.c is each independently selected from F, OH, NH.sub.2,
CN, CH.sub.3, COOH and --SO.sub.2CH.sub.3;
[0016] R is each independently selected from F, OH, NH.sub.2 and
CH.sub.3.
[0017] The present disclosure provides a compound represented by
formula (II), an isomer thereof or a pharmaceutically acceptable
salt thereof,
##STR00006##
[0018] wherein,
[0019] R.sub.1 is selected from H, F, Cl, Br, I, OH, NH.sub.2, CN,
C.sub.1-6 alkyl and cyclopropyl, wherein the C.sub.1-6 alkyl and
cyclopropyl are optionally substituted by 1, 2 or 3 R.sub.a;
[0020] R.sub.2 is selected from thienyl, phenyl, pyridyl,
cyclopropyl, cyclohexyl and
##STR00007##
wherein the thienyl, phenyl, pyridyl, cyclopropyl, cyclohexyl
and
##STR00008##
are optionally substituted by 1, 2, 3, 4 or 5 R.sub.b;
[0021] T.sub.1 is selected from CH.sub.2, NH and O;
[0022] R.sub.3 is selected from C.sub.1-6 alkyl, wherein the
C.sub.1-6 alkyl is optionally substituted by 1, 2 or 3 R.sub.c;
[0023] R.sub.a is each independently selected from F, OH, NH.sub.2
and CN;
[0024] R.sub.b is each independently selected from H, D, F, Cl, Br,
I, OH, NH.sub.2, CN, C.sub.1-3 alkyl, --C(.dbd.O)--C.sub.1-3 alkyl
and --C(.dbd.O)--O--C.sub.1-3 alkyl, wherein the OH, NH.sub.2,
C.sub.1-3 alkyl, --C(.dbd.O)--C.sub.1-3 alkyl and
--C(.dbd.O)--O--C.sub.1-3 alkyl are optionally substituted by 1, 2
or 3 R;
[0025] R.sub.c is each independently selected from F, OH, NH.sub.2,
CN, CH.sub.3, COOH and --SO.sub.2CH.sub.3;
[0026] R is each independently selected from F, OH, NH.sub.2 and
CH.sub.3.
[0027] In some embodiments of the present disclosure, the R.sub.1
is selected from H, F, Cl, Br, I, OH, NH.sub.2, CN, C.sub.1-3
alkyl, cyclopropyl and --C(.dbd.O)--NH.sub.2, wherein the C.sub.1-3
alkyl, cyclopropyl and --C(.dbd.O)--NH.sub.2 are optionally
substituted by 1, 2 or 3 R.sub.a, and the other variables are as
defined in the present disclosure.
[0028] In some embodiments of the present disclosure, the R.sub.1
is selected from H, F, Cl, Br, I, OH, NH.sub.2, CN, C.sub.1-3 alkyl
and cyclopropyl, wherein the C.sub.1-3 alkyl and cyclopropyl are
optionally substituted by 1, 2 or 3 R.sub.a, and the other
variables are as defined in the present disclosure.
[0029] In some embodiments of the present disclosure, the R.sub.1
is selected from CN, CH.sub.3, CF.sub.3,
##STR00009##
and --C(.dbd.O)--NH.sub.2, and the other variables are as defined
in the present disclosure.
[0030] In some embodiments of the present disclosure, the R.sub.1
is selected from CN, CH.sub.3, CF.sub.3,
##STR00010##
and the other variables are as defined in the present
disclosure.
[0031] In some embodiments of the present disclosure, the R.sub.b
is each independently selected from H, D, F, Cl, Br, I, OH,
NH.sub.2, CN, CH.sub.3, CH.sub.2CH.sub.3, CH.sub.2CH.sub.2CH.sub.3,
CH(CH.sub.3).sub.2, COOH,
##STR00011##
wherein the OH, NH.sub.2, CH.sub.3, CH.sub.2CH.sub.3,
CH.sub.2CH.sub.2CH.sub.3, CH(CH.sub.3).sub.2,
##STR00012##
are optionally substituted by 1, 2 or 3 R, and the other variables
are as defined in the present disclosure.
[0032] In some embodiments of the present disclosure, the R.sub.b
is each independently selected from H, D, F, Cl, Br, I, OH,
NH.sub.2, CN, CH.sub.3, CH.sub.2CH.sub.3, CH.sub.2CH.sub.2CH.sub.3,
CH(CH.sub.3).sub.2, COOH,
##STR00013##
wherein the OH, NH.sub.2, CH.sub.3, CH.sub.2CH.sub.3,
CH.sub.2CH.sub.2CH.sub.3, CH(CH.sub.3).sub.2,
##STR00014##
are optionally substituted by 1, 2 or 3 R, and the other variables
are as defined in the present disclosure.
[0033] In some embodiments of the present disclosure, the R.sub.b
is each independently selected from H, D, F, Cl, OH, OCH.sub.3, CN,
CH.sub.3, CH.sub.2OH, CH.sub.2NH.sub.2, COOH,
##STR00015##
and the other variables are as defined in the present
disclosure.
[0034] In some embodiments of the present disclosure, the R.sub.2
is selected from thienyl, phenyl, pyridyl, cyclopropyl,
cyclohexyl
##STR00016##
wherein the thienyl, phenyl, pyridyl, cyclopropyl, cyclohexyl,
##STR00017##
are optionally substituted by 1, 2, 3, 4 or 5 R.sub.b, and the
other variables are as defined in the present disclosure.
[0035] In some embodiments of the present disclosure, the R.sub.2
is selected from
##STR00018##
and the other variables are as defined in the present
disclosure.
[0036] In some embodiments of the present disclosure, the R.sub.2
is selected from
##STR00019##
and the other variables are as defined in the present
disclosure.
[0037] In some embodiments of the present disclosure, the R.sub.2
is selected from
##STR00020##
and the other variables are as defined in the present
disclosure.
[0038] In some embodiments of the present disclosure, the R.sub.3
is selected from C.sub.2-5 alkyl, wherein the C.sub.2-5 alkyl is
optionally substituted by 1, 2 or 3 R.sub.c, and the other
variables are as defined in the present disclosure.
[0039] In some embodiments of the present disclosure, the R.sub.3
is selected from CH.sub.2CH.sub.3, CH.sub.2CH.sub.2CH.sub.3,
CH(CH.sub.3).sub.2, CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
CH.sub.2CH(CH.sub.3).sub.2 and CH.sub.2CH.sub.2CH(CH.sub.3).sub.2,
wherein the CH.sub.2CH.sub.3, CH.sub.2CH.sub.2CH.sub.3,
CH(CH.sub.3).sub.2, CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
CH.sub.2CH(CH.sub.3).sub.2 and CH.sub.2CH.sub.2CH(CH.sub.3).sub.2
are optionally substituted by 1, 2 or 3 R.sub.c, and the other
variables are as defined in the present disclosure.
[0040] In some embodiments of the present disclosure, the R.sub.3
is selected from
##STR00021##
and the other variables are as defined in the present
disclosure.
[0041] The present disclosure also provides a compound represented
by formula (II), an isomer thereof or a pharmaceutically acceptable
salt thereof,
##STR00022##
[0042] wherein,
[0043] R.sub.1 is selected from H, F, Cl, Br, I, OH, NH.sub.2, CN,
C.sub.1-6 alkyl and cyclopropyl, wherein the C.sub.1-6 alkyl and
cyclopropyl are optionally substituted by 1, 2 or 3 R.sub.a;
[0044] R.sub.2 is selected from phenyl, pyridyl, cyclopropyl,
cyclohexyl and
##STR00023##
wherein the phenyl, pyridyl, cyclopropyl, cyclohexyl and
##STR00024##
are optionally substituted by 1, 2 or 3 R.sub.b;
[0045] T.sub.1 is selected from CH.sub.2, NH and O;
[0046] R.sub.3 is selected from C.sub.1-6 alkyl, wherein the
C.sub.1-6 alkyl is optionally substituted by 1, 2 or 3 R.sub.c;
[0047] R.sub.a is each independently selected from F, OH, NH.sub.2
and CN;
[0048] R.sub.b is each independently selected from H, F, OH,
NH.sub.2, CN, CH.sub.3, --C(.dbd.O)--C.sub.1-3 alkyl and
--C(.dbd.O)--O--C.sub.1-3 alkyl, wherein the CH.sub.3,
--C(.dbd.O)--C.sub.1-3 alkyl and --C(.dbd.O)--O--C.sub.1-3 alkyl
are optionally substituted by 1, 2 or 3 R;
[0049] R.sub.c is each independently selected from F, OH, NH.sub.2,
CN, CH.sub.3 and C(.dbd.O);
[0050] R is each independently selected from F, OH and
NH.sub.2.
[0051] In some embodiments of the present disclosure, the R.sub.1
is selected from H, F, Cl, Br, I, OH, NH.sub.2, CN, C.sub.1-3 alkyl
and cyclopropyl, wherein the C.sub.1-3 alkyl and cyclopropyl are
optionally substituted by 1, 2 or 3 R.sub.a, and the other
variables are as defined in the present disclosure.
[0052] In some embodiments of the present disclosure, the R.sub.1
is selected from CN, CH.sub.3, CF.sub.3,
##STR00025##
and the other variables are as defined in the present
disclosure.
[0053] In some embodiments of the present disclosure, the R.sub.b
is independently each selected from H, F, Cl, Br, I, OH, NH.sub.2,
CN, CH.sub.3,
##STR00026##
wherein the CH.sub.3,
##STR00027##
are optionally substituted by R, and the other variables are as
defined in the present disclosure.
[0054] In some embodiments of the present disclosure, the R.sub.b
is each independently selected from H, F, OH, CN, CH.sub.3,
CH.sub.2OH, CH.sub.2NH.sub.2,
##STR00028##
and the other variables are as defined in the present
disclosure.
[0055] In some embodiments of the present disclosure, the R.sub.2
is selected from phenyl, pyridyl, cyclopropyl, cyclohexyl,
##STR00029##
wherein the phenyl, pyridyl, cyclopropyl, cyclohexyl,
##STR00030##
are optionally substituted by 1, 2 or 3 R.sub.b, and the other
variables are as defined in the present disclosure.
[0056] In some embodiments of the present disclosure, the R.sub.2
is selected from
##STR00031##
and the other variables are as defined in the present
disclosure.
[0057] In some embodiments of the present disclosure, the R.sub.2
is selected from
##STR00032##
and the other variables are as defined in the present
disclosure.
[0058] In some embodiments of the present disclosure, the R.sub.3
is selected from C.sub.2-5 alkyl, wherein the C.sub.2-5 alkyl is
optionally substituted by 1, 2 or 3 R.sub.c, and the other
variables are as defined in the present disclosure.
[0059] In some embodiments of the present disclosure, the R.sub.3
is selected from
##STR00033##
and the other variables are as defined in the present
disclosure.
[0060] The present disclosure also provides a compound represented
by formula (I), an isomer thereof or a pharmaceutically acceptable
salt thereof,
##STR00034##
[0061] wherein,
[0062] R.sub.1 is selected from H, F, Cl, Br, I, OH, NH.sub.2, CN
and C.sub.1-6 alkyl, wherein the C.sub.1-6 alkyl is optionally
substituted by 1, 2 or 3 R.sub.a;
[0063] R.sub.2 is selected from C.sub.3-8 cycloalkyl and 3-8
membered heterocycloalkyl, wherein the C.sub.3-8 cycloalkyl and 3-8
membered heterocycloalkyl are optionally substituted by 1, 2 or 3
R.sub.b;
[0064] L.sub.1 is selected from C.sub.1-6 alkyl, wherein the
C.sub.1-6 alkyl is optionally substituted by 1, 2 or 3 R.sub.c;
[0065] R.sub.a is each independently selected from F, Cl, Br, I,
OH, NH.sub.2 and CN;
[0066] R.sub.b is each independently selected from H, F, Cl, Br, I,
OH, NH.sub.2, CN, CH.sub.3, --C(.dbd.O)--C.sub.1-3 alkyl and
--C(.dbd.O)--C.sub.1-3 alkoxy;
[0067] R.sub.c is each independently selected from F, Cl, Br, I,
OH, NH.sub.2, CN and CH.sub.3;
[0068] the 3-8 membered heterocycloalkyl contains 1, 2 or 3
heteroatoms or heteroatom groups independently selected from
--NH--, N and O.
[0069] In some embodiments of the present disclosure, the R.sub.1
is selected from H, F, Cl, Br, I, OH, NH.sub.2, CN and C.sub.1-3
alkyl, wherein the C.sub.1-3 alkyl is optionally substituted by 1,
2 or 3 R.sub.a, and the other variables are as defined in the
present disclosure.
[0070] In some embodiments of the present disclosure, the R.sub.1
is CH.sub.3, and the other variables are as defined in the present
disclosure.
[0071] In some embodiments of the present disclosure, the R.sub.b
is selected from H, F, Cl, Br, I, OH, NH.sub.2, CN, CH.sub.3,
##STR00035##
and the other variables are as defined in the present
disclosure.
[0072] In some embodiments of the present disclosure, the R.sub.2
is selected from C.sub.3-6 cycloalkyl and 4-6 membered
heterocycloalkyl, wherein the C.sub.3-6 cycloalkyl and 4-6 membered
heterocycloalkyl are optionally substituted by 1, 2 or 3 R.sub.b,
and the other variables are as defined in the present
disclosure.
[0073] In some embodiments of the present disclosure, the R.sub.2
is selected from morpholinyl, piperidinyl, piperazinyl,
tetrahydropyranyl and cyclopropyl, wherein the morpholinyl,
piperidinyl, piperazinyl, tetrahydropyranyl and cyclopropyl are
optionally substituted by 1, 2 or 3 R.sub.b, and the other
variables are as defined in the present disclosure.
[0074] In some embodiments of the present disclosure, the R.sub.2
is selected from
##STR00036##
and the other variables are as defined in the present
disclosure.
[0075] In some embodiments of the present disclosure, the Li is
selected from C.sub.3-5 alkyl, wherein the C.sub.3-5 alkyl is
optionally substituted by 1, 2 or 3 R.sub.c, and the other
variables are as defined in the present disclosure.
[0076] In some embodiments of the present disclosure, the L.sub.1
is
##STR00037##
and the other variables are as defined in the present
disclosure.
[0077] In some embodiments of the present disclosure, the compound,
the isomer or the pharmaceutically acceptable salt thereof, and the
compound is selected from:
##STR00038## ##STR00039##
[0078] wherein, L.sub.1 is selected from C.sub.2-5 alkyl, and
R.sub.1, R.sub.3 and R.sub.b are as defined in the present
disclosure.
[0079] There are also some embodiments of the present disclosure
obtained by an arbitrary combination of the above variables.
[0080] The present disclosure also provides a compound represented
by the following formula, an isomer thereof or a pharmaceutically
acceptable salt thereof,
##STR00040## ##STR00041## ##STR00042## ##STR00043## ##STR00044##
##STR00045## ##STR00046## ##STR00047##
[0081] The present disclosure also provides a pharmaceutical
composition, comprising a therapeutically effective amount of the
compound described above, the isomer thereof or the
pharmaceutically acceptable salt thereof as an active ingredient
and a pharmaceutically acceptable carrier.
[0082] The present disclosure also provides a use of the compound
described above, the isomer thereof or the pharmaceutically
acceptable salt thereof, or the pharmaceutical composition
described above in the preparation of a medicament for treating
diseases related to IRAK4 and BTK.
Technical Effect
[0083] The compound of the present disclosure generally exhibits
good inhibitory activity against IRAK4 and BTK. The compound of the
present disclosure generally exhibits a good activity of inhibiting
cell TNF-.alpha. production in THP-1 cells, a good activity of
inhibiting cell proliferation in OCI-LY10, OCI-LY3 and TMD-8 cells,
and a good in vivo efficacy in subcutaneous xenograft tumor model
of human B-cell lymphoma OCI-LY10 cells.
Definition and Description
[0084] Unless otherwise specified, the following terms and phrases
when used herein have the following meanings. A specific term or
phrase should not be considered indefinite or unclear in the
absence of a particular definition, but should be understood in the
ordinary sense. When a trading name appears herein, it is intended
to refer to its corresponding commodity or active ingredient
thereof.
[0085] The term "pharmaceutically acceptable" is used herein in
terms of those compounds, materials, compositions, and/or dosage
forms, which are suitable for use in contact with human and animal
tissues within the scope of reliable medical judgment, with no
excessive toxicity, irritation, an allergic reaction or other
problems or complications, commensurate with a reasonable
benefit/risk ratio.
[0086] The term "pharmaceutically acceptable salt" refers to a salt
of the compound of the present disclosure that is prepared by
reacting the compound having a specific substituent of the present
disclosure with a relatively non-toxic acid or base. When the
compound of the present disclosure contains a relatively acidic
functional group, a base addition salt can be obtained by bringing
the neutral form of the compound into contact with a sufficient
amount of base in a pure solution or a suitable inert solvent. The
pharmaceutically acceptable base addition salt includes a salt of
sodium, potassium, calcium, ammonium, organic amine or magnesium,
or similar salts. When the compound of the present disclosure
contains a relatively basic functional group, an acid addition salt
can be obtained by bringing the neutral form of the compound into
contact with a sufficient amount of acid in a pure solution or a
suitable inert solvent. Examples of the pharmaceutically acceptable
acid addition salt include an inorganic acid salt, wherein the
inorganic acid includes, for example, hydrochloric acid,
hydrobromic acid, nitric acid, carbonic acid, bicarbonate,
phosphoric acid, monohydrogen phosphate, dihydrogen phosphate,
sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid,
and the like; and an organic acid salt, wherein the organic acid
includes, for example, acetic acid, propionic acid, isobutyric
acid, maleic acid, malonic acid, benzoic acid, succinic acid,
suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic
acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid,
tartaric acid, and methanesulfonic acid, and the like; and salts of
amino acid (such as arginine and the like), and a salt of an
organic acid such as glucuronic acid and the like. Certain specific
compounds of the present disclosure contain both basic and acidic
functional groups, thus can be converted to any base or acid
addition salt.
[0087] The pharmaceutically acceptable salt of the present
disclosure can be prepared from the parent compound that contains
an acidic or basic moiety by conventional chemical method.
Generally, such salt can be prepared by reacting the free acid or
base form of the compound with a stoichiometric amount of an
appropriate base or acid in water or an organic solvent or a
mixture thereof.
[0088] The compounds of the present disclosure may exist in
specific geometric or stereoisomeric forms. The present disclosure
contemplates all such compounds, including cis and trans isomers,
(-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers
isomers, (D)-isomers, (L)-isomers, and racemic and other mixtures
thereof, such as enantiomers or diastereomeric enriched mixtures,
all of which are within the scope of the present disclosure.
Additional asymmetric carbon atoms may be present in substituents
such as alkyl. All these isomers and their mixtures are included
within the scope of the present disclosure.
[0089] Unless otherwise specified, D in the present disclosure
represents deuterium (.sup.2H).
[0090] Unless otherwise specified, the term "enantiomer" or
"optical isomer" refers to stereoisomers that are mirror images of
each other.
[0091] Unless otherwise specified, the term "cis-trans isomer" or
"geometric isomer" is caused by the inability to rotate freely of
double bonds or single bonds of ring-forming carbon atoms.
[0092] Unless otherwise specified, the term "diastereomer" refers
to a stereoisomer in which a molecule has two or more chiral
centers and the relationship between the molecules is not mirror
images.
[0093] Unless otherwise specified, "(+)" refers to dextrorotation,
"(-)" refers to levorotation, and or "(.+-.)" refers to
racemic.
[0094] Unless otherwise specified, the absolute configuration of a
stereogenic center is represented by a wedged solid bond () and a
wedged dashed bond (), and the relative configuration of a
stereogenic center is represented by a straight solid bond () and a
straight dashed bond (), a wave line () is used to represent a
wedged dashed bond () or a wedged dashed bond (), or the wave line
() is used to represent a straight solid bond () and a straight
dashed bond ().
[0095] Unless otherwise specified, when double bond structure, such
as carbon-carbon double bond, carbon-nitrogen double bond, and
nitrogen-nitrogen double bond, exists in the compound, and each of
the atoms on the double bond is connected to two different
substituents (including the condition where a double bond contains
a nitrogen atom, the lone pair of electrons attached on the
nitrogen atom is regarded as a substituent connected), if the atom
on the double bond in the compound is connected to its substituent
by a wave line (), this refers to the (Z) isomer, (E) isomer or a
mixture of two isomers of the compound. For example, the following
formula (A) means that the compound exists as a single isomer of
formula (A-1) or formula (A-2) or as a mixture of two isomers of
formula (A-1) and formula (A-2); the following formula (B) means
that the compound exists in the form of a single isomer of formula
(B-1) or formula (B-2) or in the form of a mixture of two isomers
of formula (B-1) and formula (B-2). The following formula (C) means
that the compound exists as a single isomer of formula (C-1) or
formula (C-2) or as two a mixture of two isomers of formula (C-1)
and formula (C-2).
##STR00048##
[0096] Unless otherwise specified, the term "tautomer" or
"tautomeric form" means that at room temperature, the isomers of
different functional groups are in dynamic equilibrium and can be
transformed into each other quickly. If tautomers possibly exist
(such as in solution), the chemical equilibrium of tautomers can be
reached. For example, proton tautomer (also called prototropic
tautomer) includes interconversion through proton migration, such
as keto-enol isomerization and imine-enamine isomerization. Valence
tautomer includes some recombination of bonding electrons for
mutual transformation. A specific example of keto-enol
tautomerization is the tautomerism between two tautomers of
pentane-2, 4-dione and 4-hydroxypent-3-en-2-one.
[0097] Unless otherwise specified, the terms "enriched in one
isomer", "enriched in isomers", "enriched in one enantiomer" or
"enriched in enantiomers" refer to the content of one of the
isomers or enantiomers is less than 100%, and the content of the
isomer or enantiomer is greater than or equal to 60%, or greater
than or equal to 70%, or greater than or equal to 80%, or greater
than or equal to 90%, or greater than or equal to 95%, or greater
than or equal to 96%, or greater than or equal to 97%, or greater
than or equal to 98%, or greater than or equal to 99%, or greater
than or equal to 99.5%, or greater than or equal to 99.6%, or
greater than or equal to 99.7%, or greater than or equal to 99.8%,
or greater than or equal to 99.9%.
[0098] Unless otherwise specified, the term "isomer excess" or
"enantiomeric excess" refers to the difference between the relative
percentages of two isomers or two enantiomers. For example, if the
content of one isomer or enantiomer is 90%, and the content of the
other isomer or enantiomer is 10%, the isomer or enantiomer excess
(ee value) is 80%.
[0099] Optically active (R)- and (S)-isomer, or D and L isomer can
be prepared using chiral synthesis or chiral reagents or other
conventional techniques. If one kind of enantiomer of certain
compound of the present disclosure is to be obtained, the pure
desired enantiomer can be obtained by asymmetric synthesis or
derivative action of chiral auxiliary followed by separating the
resulting diastereomeric mixture and cleaving the auxiliary group.
Alternatively, when the molecule contains a basic functional group
(such as amino) or an acidic functional group (such as carboxyl),
the compound reacts with an appropriate optically active acid or
base to form a salt of the diastereomeric isomer which is then
subjected to diastereomeric resolution through the conventional
method in the art to obtain the pure enantiomer. In addition, the
enantiomer and the diastereoisomer are generally isolated through
chromatography which uses a chiral stationary phase and optionally
combines with a chemical derivative method (such as carbamate
generated from amine).
[0100] The compound of the present disclosure may contain an
unnatural proportion of atomic isotope at one or more than one
atom(s) that constitute the compound. For example, the compound can
be radiolabeled with a radioactive isotope, such as tritium
(.sup.3H), iodine-125 (.sup.125I) or C-14 (.sup.14C). For another
example, deuterated drugs can be formed by replacing hydrogen with
heavy hydrogen, and the bond formed by deuterium and carbon is
stronger than that of ordinary hydrogen and carbon; compared with
non-deuterated drugs, deuterated drugs have the advantages of
reduced toxic and side effects, increased drug stability, enhanced
efficacy, extended biological half-life of drugs, etc. All isotopic
variations of the compound of the present disclosure, whether
radioactive or not, are encompassed within the scope of the present
disclosure.
[0101] The term "optional" or "optionally" means that the
subsequent event or condition may occur but not requisite, that the
term includes the instance in which the event or condition occurs
and the instance in which the event or condition does not
occur.
[0102] The term "substituted" means one or more than one hydrogen
atom (s) on a specific atom are substituted with the substituent,
including deuterium and hydrogen variables, as long as the valence
of the specific atom is normal and the substituted compound is
stable. When the substituent is an oxygen (i.e., .dbd.O), it means
two hydrogen atoms are substituted. Positions on an aromatic ring
cannot be substituted with a ketone. The term "optionally
substituted" means an atom can be substituted with a substituent or
not, unless otherwise specified, the type and number of the
substituent may be arbitrary as long as being chemically
achievable.
[0103] When any variable (such as R) occurs in the constitution or
structure of the compound more than once, the definition of the
variable at each occurrence is independent. Thus, for example, if a
group is substituted with 0-2 R, the group can be optionally
substituted with up to two R, wherein the definition of R at each
occurrence is independent. Moreover, a combination of the
substituent and/or the variant thereof is allowed only when the
combination results in a stable compound.
[0104] When the number of a linking group is 0, such as
--(CRR).sub.0--, it means that the linking group is a single
bond.
[0105] When a substituent is vacant, it means that the substituent
does not exist, for example, when X is vacant in A-X, the structure
of A-X is actually A. When the enumerative substituent does not
indicate by which atom it is linked to the group to be substituted,
such substituent can be bonded by any atom thereof. For example,
when pyridyl acts as a substituent, it can be linked to the group
to be substituted by any carbon atom on the pyridine ring.
[0106] When the enumerative linking group does not indicate the
direction for linking, the direction for linking is arbitrary, for
example, the linking group L contained in
##STR00049##
is -M-W--, then -M-W-- can link ring A and ring B to form
##STR00050##
in the direction same as left-to-right reading order, and form
##STR00051##
in the direction contrary to left-to-right reading order. A
combination of the linking groups, substituents and/or variables
thereof is allowed only when such combination can result in a
stable compound.
[0107] Unless otherwise specified, when a group has one or more
linkable sites, any one or more sites of the group can be linked to
other groups through chemical bonds. When the linking site of the
chemical bond is not positioned, and there is H atom at the
linkable site, then the number of H atom at the site will decrease
correspondingly with the number of chemical bond linking thereto so
as to meet the corresponding valence. The chemical bond between the
site and other groups can be represented by a straight solid bond
(), a straight dashed bond () or a wavy line
##STR00052##
For example, the straight solid bond in --OCH.sub.3 means that it
is linked to other groups through the oxygen atom in the group; the
straight dashed bonds in
##STR00053##
means that it is linked to other groups through the two ends of
nitrogen atom in the group; the wave lines in
##STR00054##
means that the phenyl group is linked to other groups through
carbon atoms at position 1 and position 2;
##STR00055##
means that it can be linked to other groups through any linkable
sites on the piperidinyl by one chemical bond, including at least
four types of linkage, including
##STR00056##
Even though the H atom is drawn on the --N--,
##STR00057##
still includes the linkage of
##STR00058##
merely when one chemical bond was connected, the H of this site
will be reduced by one to the corresponding monovalent
piperidinyl.
[0108] Unless otherwise specified, the term "C.sub.1-6 alkyl"
refers to a linear or branched saturated hydrocarbon group
consisting of 1 to 6 carbon atoms. The C.sub.1-6 alkyl includes
C.sub.1-5, C.sub.1-4, C.sub.1-3, C.sub.1-2, C.sub.2-6, C.sub.2-4,
C.sub.6 and C.sub.5 alkyl and the like. It can be monovalent (such
as methyl), divalent (such as methylene) or multivalent (such as
methine). Examples of C.sub.1-6 alkyl include but are not limited
to methyl (Me), ethyl (Et), propyl (including n-propyl and
isopropyl), butyl (including n-butyl, isobutyl, s-butyl, and
t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl),
hexyl and the like.
[0109] Unless otherwise specified, the term "C.sub.2-5 alkyl"
refers to a linear or branched saturated hydrocarbon group
consisting of 2 to 5 carbon atoms. The C.sub.2-5 alkyl includes
C.sub.2-5, C.sub.2-4, C.sub.2-3, C.sub.2, C.sub.3, C.sub.4 and
C.sub.5 alkyl and the like. Examples of C.sub.2-5 alkyl include but
are not limited to ethyl (Et), propyl (including n-propyl and
isopropyl), butyl (including n-butyl, isobutyl, s-butyl, and
t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl) and
the like.
[0110] Unless otherwise specified, the term "C.sub.1-3 alkyl"
refers to a linear or branched saturated hydrocarbon group
consisting of 1 to 3 carbon atoms. The C.sub.1-3 alkyl group
includes C.sub.1-2 and C.sub.2-3 alkyl and the like; it can be
monovalent (such as methyl), divalent (such as methylene) or
multivalent (such as methine). Examples of C.sub.1-3 alkyl include
but are not limited to methyl (Me), ethyl (Et), propyl (including
n-propyl and isopropyl), etc.
[0111] Unless otherwise specified, the term "C.sub.1-3 alkoxy"
refers to an alkyl containing 1 to 3 carbon atoms that are
connected to the rest of the molecule through an oxygen atom. The
C.sub.1-3 alkoxy includes C.sub.1-2, C.sub.2-3, C.sub.1, C.sub.2
and C.sub.3 alkoxy and the like. Examples of C.sub.1-3 alkoxy
include, but are not limited to, methoxy, ethoxy, propoxy
(including n-propoxy and isopropoxy), etc.
[0112] Unless otherwise specified, "C.sub.3-8 cycloalkyl" refers to
a saturated cyclic hydrocarbon group consisting of 3 to 8 carbon
atoms, including monocyclic and bicyclic systems, wherein the
bicyclic systems include spiro ring, fused ring and bridged ring.
The C.sub.3-8 cycloalkyl includes C.sub.3-6, C.sub.3-5, C.sub.4-8,
C.sub.4-6, C.sub.4-5, C.sub.5-8 or C.sub.5-6 cycloalkyl and the
like, the C.sub.3-8 cycloalkyl can be monovalent, divalent or
multivalent. Examples of C.sub.3-8 cycloalkyl include, but are not
limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, norbornyl, [2.2.2] dicyclooctyl and the like.
[0113] Unless otherwise specified, the term "3-8 membered
heterocycloalkyl" by itself or in combination with other terms
refers to a saturated cyclic group consisting of 3 to 8 ring atoms,
wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently
selected from O, S and N, and the rest are carbon atoms, wherein
nitrogen atoms are optionally quaternized, and nitrogen and sulfur
heteroatoms can be optionally oxidized (i.e., NO and S(O).sub.p, p
is 1 or 2). The 3-8 membered heterocycloalkyl includes monocyclic
and bicyclic systems, wherein the bicyclic systems include spiro
ring, fused ring and bridged ring. In addition, with regard to the
"3-8 membered heterocycloalkyl", a heteroatom may occupy the
connection position of the heterocycloalkyl with the rest of the
molecule. The 3-8 membered heterocycloalkyl includes 3-6 membered,
3-5 membered, 4-6 membered, 5-6 membered, 4-membered, 5-membered,
and 6-membered heterocycloalkyl and the like. Examples of 3-8
membered heterocycloalkyl include, but are not limited to,
azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl,
imidazolidinyl, tetrahydrothienyl (including
tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl and the like),
tetrahydrofuranyl (including tetrahydrofuran-2-yl and the like),
tetrahydropyranyl, piperidinyl (including 1-piperidinyl,
2-piperidinyl and 3-piperidinyl and the like), piperazinyl
(including 1-piperazinyl and 2-piperazinyl and the like),
morpholinyl (including 3-morpholinyl and 4-morpholinyl and the
like), dioxinyl, dithianyl, isoxazolidinyl, isothiazolidinyl,
1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl,
homopiperidinyl or dioxacycloheptyl and the like.
[0114] The term "leaving group" refers to a functional group or
atom which can be replaced by another functional group or atom
through a substitution reaction (such as affinity substitution
reaction). For example, representative leaving groups include
triflate; chlorine, bromine, and iodine; sulfonate group, such as
mesylate, tosylate, p-bromobenzenesulfonate, p-toluenesulfonates
and the like; acyloxy, such as acetoxy, trifluoroacetoxy and the
like.
[0115] The term "protecting group" includes, but is not limited to
"amino protecting group", "hydroxy protecting group" or "thio
protecting group". The term "amino protecting group" refers to a
protecting group suitable for blocking the side reaction on the
nitrogen of an amino. Representative amino protecting groups
include, but are not limited to: formyl; acyl, such as alkanoyl
(e.g., acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl,
such as tert-butoxycarbonyl (Boc); arylmethoxycarbonyl such as
benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc);
arylmethyl, such as benzyl (Bn), trityl (Tr),
1,1-bis-(4'-methoxyphenyl)methyl; silyl, such as trimethylsilyl
(TMS) and tert-butyldimethylsilyl (TBS) and the like. The term
"hydroxy protecting group" refers to a protecting group suitable
for blocking the side reaction on hydroxy. Representative hydroxy
protecting groups include, but are not limited to: alkyl, such as
methyl, ethyl, and tert-butyl; acyl, such as chain alkanoyl (e.g.,
acetyl); arylmethyl, such as benzyl (Bn), p-methoxybenzyl (PMB),
9-fluorenylmethyl (Fm), and diphenylmethyl (benzhydryl, DPM);
silyl, such as trimethylsilyl (TMS) and tert-butyl dimethyl silyl
(TBS) and the like.
[0116] The compounds of the present disclosure can be prepared by a
variety of synthetic methods known to those skilled in the art,
including the specific embodiments listed below, the embodiments
formed by their combination with other chemical synthesis methods,
and equivalent alternatives known to those skilled in the art,
preferred implementations include but are not limited to the
embodiments of the present disclosure.
[0117] The solvent used in the present disclosure is commercially
available. The following abbreviations are used in the present
disclosure: DMSO refers to dimethyl sulfoxide; EtOH refers to
ethanol; MeOH refers to methanol; M refers to mol/L; N/A refers to
not tested; MgCl.sub.2 refers to magnesium chloride; EGTA refers to
ethylenebis(oxyethylenenitrilo)tetraacetic acid; and
Na.sub.3VO.sub.4 refers to sodium vanadate.
[0118] The compounds of the present disclosure are named according
to the conventional naming principles in the art or by
ChemDraw.RTM. software, and the commercially available compounds
use the supplier catalog names.
DESCRIPTION OF THE DRAWINGS
[0119] FIG. 1 shows a graph of plasma TNF-.alpha.
concentration.
[0120] FIG. 2 is a graph showing the weight change of mice in
different groups.
[0121] FIG. 3 is a graph of relative weight change (%).
[0122] FIG. 4 is a graph of the tumor growth curve.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0123] The following embodiment further illustrates the present
disclosure, but the present disclosure is not limited thereto. The
present disclosure has been described in detail herein, and its
specific embodiments have also been disclosed; for one skilled in
the art, it is obvious to make various modifications and
improvements to the embodiments of the present disclosure without
departing from the spirit and scope of the present disclosure.
Reference Embodiment 1: Synthesis of Intermediate A1
##STR00059##
[0125] Synthetic Route:
##STR00060##
[0126] Step 1: Synthesis of Compound A1
[0127] Ethyl succinyl chloride (50.0 g) was added to acetonitrile
(500.0 mL) and the mixture was stirred evenly;
(trimethylsilyl)diazomethane (2 M, 227.8 mL) was added dropwise to
the reaction system and the mixture was stirred at 25.degree. C.
for 0.5 hours. Then, hydrobromic acid acetic acid solution (93.1 g,
33% content) was added dropwise to the reaction system at 0.degree.
C., and the mixture was stirred at 25.degree. C. for 0.5 hours. The
reaction mixture was concentrated under reduced pressure to remove
acetonitrile; the residue was poured into ethyl acetate (500.0 mL),
and washed with saturated sodium bicarbonate aqueous solution (100
mL.times.3). The organic phase was separated and dried over an
appropriate amount of anhydrous sodium sulfate. The desiccant was
removed by filtration, and the filtrate was concentrated to dryness
under reduced pressure to obtain a crude product. The crude product
was purified by column chromatography (eluent: petroleum
ether-petroleum ether:ethyl acetate=10:1) to obtain compound
A1.
Reference Embodiment 2: Synthesis of Intermediate A2
##STR00061##
[0129] Synthetic Route:
##STR00062##
[0130] Step 1: Synthesis of Compound A2-1
[0131] Palladium acetate (2.2 g), cesium carbonate (32.3 g) and
tri-o-tolylphosphine (6.0 g) were added to a mixed solution of
4-bromo-2-methylpyridine (8.5 g), ethyl oxazole-4-carboxylate (7.0
g) and N,N-dimethylformamide (70.0 mL). The mixture was replaced
with nitrogen three times, and stirred at 100.degree. C. for 16
hours. Then the reaction mixture was cooled to room temperature,
and filtered by celite. The filtrate was concentrated to dryness
under reduced pressure to obtain a crude product. The crude product
was purified by column chromatography (eluent: petroleum
ether:ethyl acetate=10:1-0:1) to obtain compound A2-1.
[0132] Step 2: Synthesis of Compound A2
[0133] Compound A2-1 (6.5 g) was dissolved in methanol (35.0 mL)
and water (35.0 mL) and the mixture was stirred evenly; sodium
hydroxide (2.2 g) was added to the reaction system and the mixture
was stirred at 15.degree. C. for 2 hours. Methanol was removed by
concentration under reduced pressure, and the aqueous phase was
extracted with tert-butyl methyl ether (10.0 mL.times.1). The
aqueous phase was separated, and the pH value was adjusted to 3
with 1 M hydrochloric acid. The aqueous phase was concentrated to
dryness under reduced pressure, and toluene (10.0 mL) was added to
the residue and stirred evenly. The mixture was filtered, and the
filtrate was concentrated to dryness under reduced pressure to
obtain compound A2. LCMS (ESI) m/z=205.2 [M+H].sup.+. .sup.1H NMR
(400 MHz, MeOH-d.sub.4) .delta.=8.87-8.86 (m, 2H), 8.53 (s, 1H),
8.45 (d, J=6.0 Hz, 1H), 2.89 (s, 3H).
[0134] Each fragment compound in Table 1 below was synthesized with
reference to the synthesis steps of compound A2.
TABLE-US-00001 TABLE 1 MS m/z Number Structure NMR [M + H].sup.+ A3
##STR00063## .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. = 8.91 (d, J
= 5.2 Hz, 1H), 8.73 (s, 1H), 8.41 (s, 1H), 8.26 (d, J = 5.2 Hz,
1H). 259.1 A4 ##STR00064## .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. = 8.93 (s, 1H), 8.88 (d, J = 6.8 Hz, 1H), 8.56 (s, 1H),
8.46 (m, 1H), 3.47 (m, 1H), 1.52 (d, J = 7.2 Hz, 6H). 233.2 A5
##STR00065## .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. = 8.84 (s,
1H), 8.74 (d, J = 6.0 Hz, 1H), 8.21 (d, J = 6.0 Hz, 1H), 8.19 (s,
1H), 2.48 (m, 1H), 1.53 (m, 2H), 1.37 (m, 2H). 231.1 A6
##STR00066## N/A 216.0 A7 ##STR00067## N/A 234.1
[0135] Each intermediate in Table 2 below was a commercially
available reagent.
TABLE-US-00002 TABLE 2 Number Structure CAS B1 ##STR00068##
50606-31-0 B2 ##STR00069## 13889-98-0 B3 ##STR00070## 110-91-8 B4
##STR00071## 110-89-4 B5 ##STR00072## 5382-16-1 B6 ##STR00073##
57260-71-6 B7 ##STR00074## 411235-57-9 B8 ##STR00075## 38646-68-3
B9 ##STR00076## 21987-29-1 B10 ##STR00077## 98-80-6 B11
##STR00078## 6457-49-4 B12 ##STR00079## 7144-05-0 B13 ##STR00080##
1692-25-7 B14 ##STR00081## 89490-05-1 B15 ##STR00082## 2971-79-1
B16 ##STR00083## 1993-03-9 B17 ##STR00084## 768-35-4 B18
##STR00085## 138642-62-3 B19 ##STR00086## 78495-63-3 B20
##STR00087## 162101-25-9 B21 ##STR00088## 1679-18-1 B22
##STR00089## 63503-60-6 B23 ##STR00090## 6165-68-0 B24 ##STR00091##
215527-70-1 B25 ##STR00092## 156545-07-2 B26 ##STR00093##
1256345-60-4 B27 ##STR00094## 1765-93-1
Embodiment 1: Synthesis of Compound WX001
##STR00095##
[0137] Synthetic Route:
##STR00096##
[0138] Step 1: Synthesis of Compound WX001-1
[0139] 4-Chloro-5-nitro-pyridin-2-amine (25.0 g) was dissolved in
tetrahydrofuran (200.0 mL) and then piperidine (61.3 g) was added.
The mixture was stirred at 10.degree. C. for 12 hours, the reaction
mixture was concentrated to dryness under reduced pressure; ethyl
acetate (100.0 mL) was added to the residue and the mixture was
slurried. Then the mixture was filtered, and the filtrate was
collected. The filtrate was concentrated to dryness under reduced
pressure to obtain a crude product, then the crude product was
purified by column chromatography (petroleum ether:ethyl
acetate=5:1-0:1) to obtain compound WX001-1.
[0140] Step 2: Synthesis of Compound WX001-2
[0141] A mixture of compound WX001-1 (5.0 g) and intermediate A1
(5.0 g) was replaced with nitrogen three times, and then stirred at
100.degree. C. for 12 hours. The reaction mixture was cooled to
room temperature, then poured into water (200.0 mL), and
dichloromethane (200.0 mL.times.3) was added for extraction. The
organic phases were combined and dried over an appropriate amount
of anhydrous sodium sulfate. The desiccant was removed by
filtration, and the filtrate was concentrated to dryness under
reduced pressure to obtain a crude product. The crude product was
purified by column chromatography (eluent:
dichloromethane:methanol=100:1-10:1) to obtain compound
WX001-2.
[0142] Step 3: Synthesis of Compound WX001-3
[0143] Raney nickel (3.0 g) was added to a solution of WX001-2 (3.0
g) in EtOH (50.0 mL) and the mixture was stirred under H.sub.2 (50
Psi) at 30.degree. C. for 1 hour. The mixture was filtered to
remove the catalyst, and the filtrate was concentrated to dryness
under reduced pressure to obtain compound WX001-3.
[0144] Step 4: Synthesis of Compound WX001-4
[0145] Compound WX001-3 (3.0 g), A2 (2.9 g),
O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (6.5 g) and N,N-diisopropylethylamine (3.7 g)
were added to dichloromethane (50.0 mL) and the mixture was stirred
at 20.degree. C. for 12 hours. When the reaction was completed, the
reaction mixture was poured into saturated sodium bicarbonate
aqueous solution (50.0 mL) and the mixture was stirred evenly. The
organic phase was separated and dried over an appropriate amount of
anhydrous sodium sulfate. The desiccant was removed by filtration,
and the filtrate was concentrated to dryness under reduced pressure
to obtain a crude product. The crude product was purified by column
chromatography (pure petroleum ether, petroleum ether:ethyl
acetate=1:1, ethyl acetate:methanol=10:1) to obtain compound
WX001-4.
[0146] Step 5: Synthesis of Compound WX001
[0147] Compound WX001-4 (3.3 g) was dissolved in anhydrous
tetrahydrofuran (70.0 mL) and the reaction mixture was cooled to
10.degree. C. A solution of magnesium methyl bromide (3 M, 15.4 mL)
in ether was added dropwise to the reaction system and the mixture
was stirred at 15.degree. C. for 20 minutes. The reaction mixture
was poured into saturated ammonium chloride aqueous solution (30.0
mL), and the mixture was extracted with ethyl acetate (20.0
mL.times.3). The organic phases were combined and dried over an
appropriate amount of anhydrous sodium sulfate. The desiccant was
removed by filtration, and the filtrate was concentrated to dryness
under reduced pressure to obtain a crude product. The crude product
was purified by column chromatography (pure petroleum ether,
petroleum ether:ethyl acetate=1:1, ethyl acetate:methanol=10:1),
and purified by machine purification (column: Welch Xtimate C18
250*50 mm*10 .mu.m; mobile phase: A: aqueous solution containing 10
mM NH.sub.4HCO.sub.3, B: acetonitrile; gradient: B %: 30%-55%, 10
minutes) to obtain compound WX001. LCMS (ESI) m/z=489.3
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.=9.25 (s,
1H), 8.70 (s, 1H), 8.63 (d, J=4.8 Hz, 1H), 7.93 (s, 1H), 7.83 (d,
J=4.8 Hz, 1H), 7.50 (s, 1H), 7.31 (s, 1H), 7.14 (s, 1H), 4.14-4.12
(m, 1H), 2.99-2.97 (m, 4H), 2.81-2.77 (m, 2H), 2.64 (s, 3H),
1.99-1.96 (m, 4H), 1.91-1.89 (m, 2H), 1.87 (br s, 2H), 1.27 (s,
6H).
[0148] Each of the embodiment in the following Table 3 was
synthesized with reference to the synthesis step of Embodiment 1,
except that the B4 (piperidine ring) of Step 1 in Embodiment 1 was
replaced by the corresponding B fragment of the corresponding
Fragment 1, and the synthesis step may undergo conventional
operations such as removal of Boc, hydrolysis, formation of
tertiary alcohols with esters using methyl Grignard reagents or
Suzuki coupling, etc.
TABLE-US-00003 TABLE 3 Frag- Embodi- ment Com- MS m/z ments 1 pound
Product structure NMR [M + H].sup.+ 2 B3 WX002 ##STR00097## .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. = 9.81 (s, 1H), 9.32 (s, 1H),
9.10 (s, 1H), 8.74 (d, J = 5.6 Hz, 1H), 7.86 (s, 1H), 7.78 (d, J =
5.2 Hz, 1H), 7.72 (s, 1H), 7.28 (s, 1H), 4.36 (s, 1H), 3.94-3.89
(m, 4H), 2.95-2.99 (m, 4H), 2.69-2.67 (m, 2H), 2.60 (s, 3H),
1.77-1.73 (m, 2H), 1.15 (s, 6H). 491.1 3 B6 WX003 ##STR00098##
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. = 9.44 (s, 1H), 8.90 (s,
1H), 8.81 (d, J = 6.0 Hz, 1H), 8.40 (s, 2H), 7.83 (s, 1H), 7.44 (s,
1H), 3.53 (s, 4H), 3.42 (s, 4H), 3.08-2.94 (m, 1H), 2.91-2.83 (m,
1H), 2.81 (s, 3H), 1.86-1.82 (m, 2H), 1.21 (s, 6H). 490.2 4 B1
WX004 ##STR00099## .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. = 9.61
(s, 1H), 8.94 (s, 1H), 8.80 (d, J = 6.0 Hz, 1H), 8.26 (s, 1H), 8.18
(d, J = 4.8 Hz, 1H), 7.92 (s, 1H), 7.46 (s, 1H), 3.89 (s, 4H), 3.78
(s, 3H), 3.24 (s, 4H), 2.99- 2.95 (m, 2H), 2.79 (s, 3H), 1.97-1.92
(m, 2H), 1.32 (s, 6H). 548.4 5 B2 WX005 ##STR00100## .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. = 9.41 (s, 1H), 8.77 (s, 1H), 8.64
(d, J = 4.8 Hz, 1H), 7.89 (s, 1H), 7.82 (d, J = 4.0 Hz, 1H), 7.66
(s, 1H), 7.29 (s, 1H), 3.96-3.92 (m, 4H), 3.18-3.10 (m, 4H), 2.87-
2.83 (m, 2H), 2.63 (s, 3H), 2.23 (s, 3H), 1.93- 1.88 (m, 2H), 1.30
(s, 6H). 532.1 6 B5 WX006 ##STR00101## .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. = 9.60 (m, 1H), 8.95 (s, 1H), 8.87-8.71 (m,
1H), 8.49-8.25 (m, 2H), 7.98-7.81 (m, 1H), 7.40 (s, 1H), 3.96-3.90
(m, 1H), 3.38-3.45 (m, 2H), 3.03-2.94 (m, 2H), 2.92-2.84 (m, 5H),
2.20- 2.18 (m, 2H), 2.10-1.98 (m, 2H), 1.92-1.85 (m, 2H), 1.30 (s,
6H). 505.2 7 B7 WX007 ##STR00102## .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. = 9.25 (s, 1H), 8.78 (s, 1H), 8.64 (s, 1H),
7.98 (s, 1H), 7.91 (d, J = 5.2 Hz, 1H), 7.83 (s, 1H), 7.42 (s, 1H),
2.93- 2.90 (m, 2H), 2.66 (s, 3H), 2.21-2.17 (m, 2H), 1.94-1.90 (m,
1H), 1.29 (s, 8H), 0.94-0.88 (m, 2H). 446..3 8 B8 WX008
##STR00103## .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. = 14.15-
14.08 (m, 1H), 9.65-9.61 (m, 2H), 9.21 (s, 1H), 8.76-8.73 (m, 1H),
8.13 (s, 1H), 7.95-7.83 (m, 2H), 7.45 (s, 1H), 3.10- 3.08 (m, 4H),
2.85-2.84 (m, 2H), 2.82-2.62 (m, 3H), 1.80-1.76 (m, 6H), 1.72 (s,
6H), 1.17-1.10 (m, 6H). 517.3 9 B9 WX009 ##STR00104## .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. = 9.62-9.61 (m, 1H), 8.95 (s, 1H),
8.81-8.80 (m, 1H), 8.26 (s, 1H), 8.18 (s, 1H), 7.92 (s, 1H), 7.47
(s, 1H), 3.35-3.40 (m, 4H), 2.98- 2.94 (m, 2H), 2.79 (s, 3H), 2.46
(m, 4H), 1.94- 1.90 (m, 2H), 1.31 (s, 6H). 525.2 10 A3 WX010
##STR00105## .sup.1HNMR (400 MHz, DMSO-d.sub.6) .delta. = 9.92 (s,
1H), 9.34 (s, 1H), 9.18 (s, 1H), 9.06 (d, J = 5.2 Hz, 1H),, 8.36
(s, 1H), 8.26 (d, J = 5.2 Hz, 1H), 7.70 (s, 1H), 7.22 (s, 1H),
4.25-4.20 (m, 1H), 2.93- 2.90 (m, 4H), 2.69-2.65 (m, 2H), 1.92-1.86
(m, 4H), 1.77-1.73 (m, 2H), 1.66-1.65 (m, 2H), 1.15 (s, 6H) 543.2
11 A4 WX011 ##STR00106## .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. = 9.86 (s, 1H), 9.34 (s, 1H), 9.08 (s, 1H), 8.75 (d, J =
5.2 Hz, 1H), 7.87 (s, 1H), 7.76- 7.75 (m, 1H), 7.69 (s, 1H), 7.20
(s, 1H), 4.36- 4.33 (m, 1H), 3.21-3.18 (m, 1H), 2.93-2.91 (m, 4H),
2.69-2.65 (m, 2H), 1.90-1.87 (m, 4H), 1.77- 1.69 (m, 2H), 1.75-1.58
(m, 2H), 1.36-1.25 (m, 6H), 1.14 (s, 6H). 517.3 12 A5 WX012
##STR00107## .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. = 9.87 (s,
1H), 9.35 (s, 1H), 9.09 (s, 1H), 8.67 (d, J = 4.8 Hz, 1H), 7.83 (s,
1H), 7.70- 7.68 (m, 2H), 7.21 (s, 1H), 4.36-4.33 (m, 1H), 2.93-2.92
(m, 4H), 2.90 (m, 1H) 1.92-1.87 (m, 4H), 1.77-1.74 (m, 4H), 1.23
(s, 2H), 1.15 (s, 6H), 1.08-1.06 (m, 2H), 0.99- 0.97 (m, 2H). 515.3
13 B11 WX013 ##STR00108## .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. = 9.81 (s, 1H), 9.37 (s, 1H), 9.09 (d, J = 5.2 Hz, 1H),
8.70- 8.69 (d, J = 9.2 Hz, 1H), 7.83-7.81 (m, 2H), 7.74 (s, 1H),
7.25 (s, 1H), 4.67 (m, 1H), 3.36-3.31 (m, 4H), 3.15-3.13 (m, 2H),
2.90-2.86 (m, 3H), 2.76 (m, 4H), 2.64-2.61 (m, 5H), 1.90-1.88 (m,
4H), 1.64 (m, 3H). 519.2 14 B12 WX014 ##STR00109## N/A 518.2 15 A6
WX015 ##STR00110## .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. =
9.88 (s, 1H), 9.33 (s, 1H), 9.21 (s, 1H), 9.05 (s, 1H), 8.51 (s,
1H), 8.26-8.23 (m, 1H), 7.71 (s, 1H), 7.23 (s, 1H), 4.35 (s, 1H),
2.94-2.93 (m, 7H), 2.90 (s, 3H), 2.71-2.69 (m, 2H), 2.34 (s, 1H),
1.89-1.74 (m, 2H), 1.66-1.69 (m, 2H), 1.25 (s, 3H) 500.3 44 A7
WX044 ##STR00111## .sup.1H NMR (400 MHz, DMSO-d.sub.6)= 9.95 (s,
1H), 9.35 (s, 1H), 9.14 (s, 1H), 8.91 (s, 1H), 8.70 (s, 1H), 8.26
(s, 1H), 8.13- 8.11 (m, 1H), 7.81 (s, 1H), 7.71 (s, 1H), 7.22 (s,
1H), 5.76 (s, 1H), 4.34 (s, 1H), 4.10 (s, 1H), 3.18- 3.17 (m, 2H),
2.93-2.91 (m, 3H), 1.92-1.90 (m, 3H), 1.78-1.74 (m, 4H), 1.16 (s,
6H) 518.4 45 B15 WX045 ##STR00112## N/A 547.3 46 B15 WX046
##STR00113## .sup.1H NMR (400 MHz, DMSO-d.sub.6)= 9.83 (s, 1H),
9.37 (s, 1H), 9.10 (s, 1H), 8.70 (s, 1H), 7.86- 7.83 (m, 2H), 7.72
(s, 1H), 7.22 (s, 1H), 3.17- 3.14 (m, 3H), 2.85-2.78 (m, 4H), 2.62
(s, 5H), 2.08-2.03 (m, 4H), 1.78- 1.74 (m, 2H), 1.16 (s, 6H).
533.2
Embodiment 16: Synthesis of Compound WX016
##STR00114##
[0150] Synthetic Route:
##STR00115##
[0151] WX016-1 was synthesized with reference to the synthesis step
of embodiment 16, except that the piperidine in step 1 was replaced
with 4,4-dimethylpiperidine.
[0152] Step 1: Synthesis of Compound 16
[0153] Compound WX016-1 (15.0 mg) was dissolved in a solution of
sodium hydroxide (2.3 mg) in water (1.0 mL), then methanol (1.0 mL)
was added and the reaction was carried out at 25.degree. C. for 2
hours. The pH value of the reaction mixture was adjusted to 6-7
with 1.0 M hydrochloric acid, and then the mixture was extracted
with ethyl acetate (30.0 mL.times.4), and the organic phases were
combined, dried over anhydrous sodium sulfate, filtered and
concentrated under reduced pressure. Then the residue was purified
by machine purification (column: Welch Xtimate C18 150*25 mm*5
.mu.m; mobile phase: [aqueous solution containing (10.0 mM)
NH.sub.4HCO.sub.3)-acetonitrile]; gradient: B %: 15%-50%, 10.5 min)
to obtain compound WX016. LCMS (ESI) m/z: 503.3 [M+H].sup.+.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.=9.74 (s, 1H), 9.35 (s,
1H), 9.06 (s, 1H), 8.67 (d, J=5.2 Hz, 1H), 7.83 (s, 1H), 7.76 (d,
J=5.2 Hz, 1H), 7.70-7.68 (m, 1H), 7.29 (s, 1H), 2.92-2.89 (m, 4H),
2.87-2.83 (m, 4H), 2.65-2.55 (m, 4H), 1.68 (s, 4H), 1.07 (s,
6H).
[0154] Each of the embodiments in the following Table 4 was
synthesized with reference to the synthesis step of embodiment 16,
except that the 4,4-dimethylpiperidine in step 1 was replaced with
Fragment 1.
TABLE-US-00004 TABLE 4 Embodi- Frag- MS m/z ments ment 1 Compound
Product structure NMR [M + H].sup.+ 17 B9 WX017 ##STR00116##
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. = 9.86 (s, 1H), 9.34
(s, 1H), 9.13 (s, 1 H), 8.71-8.69 (d, J = 8 Hz, 1H), 7.83 (s, 1H),
7.77-7.75 (m, 2H), 7.38 (s, 1H), 3.12 (m, 4H), 2.91-2.87 (m, 2H),
2.68-2.67 (m, 2H), 2.65-2.61 (m, 2H), 2.59 (s, 3H), 2.34 (m, 3H).
511.2 18 B4 WX018 ##STR00117## .sup.1H NMR (400 MHz, DMSO- d.sub.6)
.delta. = 9.88 (s, 1H), 9.34 (s, 1H), 9.10 (s, 1H), 8.73-8.72 (d, J
= 5.2 Hz, 1H), 7.85 (s, 1H), 7.75-7.73 (m, 2H), 7.23 (s, 1H),
2.92-2.85 (m, 6H), 2.64-2.60 (m, 6H), 1.88 (m, 4H), 1.67 (m, 2H).
475.2 19 B12 WX019 ##STR00118## N/A 504.2 20 B11 WX020 ##STR00119##
.sup.1H NMR (400 MHz, DMSO- d.sub.6) .delta. = 9.81 (s, 1H), 9.37
(s, 1H), 9.09 (m, 1H), 8.70-8.69 (d, J = 8.0 Hz, 1H), 7.83-7.81 (m,
2H), 7.74 (s, 1H), 7.25 (s, 1H), 4.67 (m, 1H), 3.15-3.13 (m, 1H),
2.90-2.86 (m, 3H), 2.76 (m, 4H), 2.64-2.61 (m, 6H), 1.90-1.88 (m,
2H), 1.64 (m, 3H). 505.2
Embodiment 21: Synthesis of Compound WX021
##STR00120##
[0156] Synthetic Route:
##STR00121##
[0157] WX021-1 was synthesized with reference to the similar
synthesis step of Embodiment 1, except that in the step 1 of
Embodiment 1, piperidine was not used to substitute the chlorine
atom.
[0158] Step 1: Synthesis of Compound WX021
[0159] WX020-1 (3.3 mg), B10 (3.33 mg), toluene (1.0 mL), ethanol
(0.5 mL) and water (0.3 mL) were added to a reaction flask, then
sodium bicarbonate (5.7 mg) and
tetrakis(triphenylphosphine)palladium (5.3 mg) were added thereto.
The mixture was replaced with nitrogen three times, and stirred at
80.degree. C. for 12 hours. Then the mixture was filtered, and the
filtrate was collected, then concentrated to dryness under reduced
pressure. The crude product was purified by silica gel plate
(eluent: dichloromethane:methanol=10:1) to obtain compound WX021.
LCMS (ESI) m/z: 482.3[M+H].sup.+. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta.=9.70 (s, 1H), 8.99 (s, 1H), 8.93 (s, 1H),
8.68-8.67 (d, J=4 Hz, 1H), 7.80 (s, 1H), 7.72 (s, 1H), 7.65-7.64
(d, J=4 Hz, 1H), 7.57-7.45 (m, 6H), 4.34 (s, 1H), 2.78-2.74 (m,
2H), 2.59 (s, 3H), 1.82-1.78 (m, 2H), 1.17 (s, 6H).
[0160] Each of the embodiment in the following Table 5 was
synthesized with reference to the synthesis step of Embodiment 21,
except that the B10 (phenylboronic acid) in the steps of Embodiment
21 was replaced by the corresponding B fragment of the
corresponding Fragment 1, and the synthesis steps may undergo
simple operations such as hydrogenation and cyanohydrolysis to
amide.
TABLE-US-00005 TABLE 5 Embodi- Frag- MS m/z ments ment 1 Compound
Product structure NMR [M + H].sup.+ 22 B14 WX022 ##STR00122##
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. = 8.72 (s, 1H),
8.65-8.64 (d, J = 4.0 Hz 1H), 8.59 (s, 1H), 8.02 (s, 1H), 7.94-7.92
(d, J = 8.0 Hz 1H), 7.59 (s, 1H), 7.37 (s, 1H), 2.87-2.79 (m, 3H),
2.67 (s, 3H), 2.00-1.90 (m, 6H), 1.80-1.77 (m, 2H), 1.54- 1.37 (m,
6H), 1.28 (s, 6H). 488.2 23 B13 WX023 ##STR00123## .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. = 10.03 (s, 1H), 8.88 (s, 1H), 8.82 (s,
1H), 8.72 (s, 1H), 8.68-8.66 (d, J = 8.0 Hz 1H), 8.57-8.56 (d, J =
4.0 Hz 1H), 7.94-7.92 (m, 1H), 7.82 (s, 1H), 7.79 (s, 1H),
7.71-7.69 (m, 1H), 7.47-7.43 (m, 1H), 2.98- 2.94 (t, J = 6.0 Hz,
2H), 2.70 (m, 4H), 2.58 (s, 3H). 469.1 24 B16 WX024 ##STR00124##
N/A 500.2 25 B17 WX025 ##STR00125## .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. = 9.83 (s, 1H), 8.92 (s, 2H), 8.67 (s, 1H),
7.81 (s, 1H), 7.77 (s, 1H), 7.70-7.68 (m, 1H), 7.53- 7.51 (m, 2H),
7.56-7.38 (m, 2H), 7.30-7.27 (m, 1H), 4.32 (s, 1H), 2.79- 2.73 (m,
2H), 2.60-2.58 (m, 3H), 1.83-1.79 (m, 2H) 1.17-1.16 (s, 6H). 500.3
26 B27 WX026 ##STR00126## .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. = 9.74 (s, 1H), 8.92 (s, 2H), 8.67 (d, J = 5.20 Hz, 1H),
7.79 (s, 1H), 7.75 (s, 1H), 7.70- 7.68 (m, 1H), 7.59-7.51 (m, 2H),
7.46 (s, 1H), 7.36- 7.40 (m, 2H), 4.32 (s, 1H), 2.79-2.73 (m, 2H),
2.60- 2.58 (m, 3H), 1.83-1.79 (m, 2H) 1.17-1.16 (s, 6H). 500.3 27
B18 WX027 ##STR00127## N/A 525.2 28 B19 WX028 ##STR00128## .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. = 9.19 (s, 1H), 9.01 (s, 2H),
9.97 (s, 1H), 8.70-8.68 (d, J = 8.0 Hz 1H), 7.88 (s, 1H), 7.65-7.57
(m, 3H), 7.46 (s, 1H), 7.17-7.15 (d, J = 8.0 Hz, 1H), 4.35 (s, 1H),
3.80 (s, 3H), 2.77-2.73 (m, 2H), 2.61 (s, 3H), 1.81- 1.77 (m, 2H),
1.16 (s, 6H). 530.3 29 B20 WX029 ##STR00129## .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. = 9.66 (s, 1H), 8.91 (d, J = 5.2 Hz, 2H),
8.67 (d, J = 5.2 Hz, 1H), 7.94-7.92 (m, 1H), 7.82 (s, 1H), 7.79 (s,
1H), 7.71-7.69 (m, 2H), 7.47-7.43 (m, 2H), 4.33 (s, 1H), 2.79-2.75
(m, 2H), 2.58 (s, 3H) 1.82-1.78 (m, 2H), 1.16 (s, 6H) 518.3 30 B21
WX030 ##STR00130## .sup.1H NMR = (400 MHz, DMSO-d.sub.6) .delta. =
9.76 (s, 1H), 8.91 (s, 2H), 8.67 (s, 1H), 7.79 (s, 1H), 7.75 (s,
1H), 7.69-7.67 (m, 1H), 7.56- 7.55 (m, 4H), 7.48 (s, 1H), 4.31 (s,
1H), 2.78-2.74 (m, 2H), 2.60-2.50 (m, 3H), 1.82-1.78 (m, 2H) 1.16-
1.15 (m, 6H) 516.3 31 B22 WX031 ##STR00131## .sup.1H NMR (400 MHz,
DMSO-d.sub.6)= 9.81 (s, 1H), 8.90 (s, 1H), 8.88 (s, 1H), 8.67 (d, J
= 8.0 Hz 1H), 7.79 (s, 1H), 7.76 (s, 1H), 7.70-7.68 (m, 1H), 7.64
(m, 1H), 7.50-7.48 (m, 4H), 4.30 (s, 1H), 2.77-2.73 (m, 2H), 2.58
(s, 3H), 1.81-1.77 (m, 2H), 1.16 (s, 6H). 516.3 32 B23 WX032
##STR00132## N/A 488.3 33 B24 WX033 ##STR00133## .sup.1H NMR (400
MHz, DMSO-d.sub.6)= 9.68 (s, 1H), 8.99 (s, 1H), 8.92 (s, 1H), 8.68
(d, J = 4.0 Hz 1H), 7.80 (s, 1H), 7.72 (s, 1H), 7.65- 7.64 (s, 1H),
7.45 (s, 1H), 4.32 (s, 1H), 2.78-2.73 (m, 2H), 2.59 (s, 3H),
1.82-1.78 (m, 2H), 1.17 (s, 6H). 487.4 34 B25 WX034 ##STR00134##
.sup.1H NMR (400 MHz, DMSO-d.sub.6)= 9.97 (s, 1H), 8.94 (s, 1H),
8.87 (s, 1H), 8.70 (s, 1H), 7.81 (s, 2H), 7.73-7.72 (m, 1H), 7.59
(s, 1H), 7.35-7.31 (m, 3H), 4.34 (s, 1H), 2.80-2.76 (m, 2H), 2.61
(s, 3H), 1.83- 1.79 (m, 2H), 1.18-1.16 (m, 6H). 518.4 35 B18 WX035
##STR00135## .sup.1H NMR (400 MHz, DMSO-d.sub.6)= 9.29 (s, 1H),
9.11 (s, 1H), 8.87-8.83 (m, 2H), 8.70 (d, J = 5.2 Hz 1H), 8.43 (m,
2H), 8.05 (s, 1H), 7.92-7.90 (m, 2H), 7.82 (s, 1H), 7.81-7.75 (m,
1H), 4.38 (s, 1H) 2.88 (t, J = 4.8 Hz, 2H), 2.60 (s, 3H), 1.87 (t,
J = 4.4 Hz, 2H), 1.19 (s, 6H) 507.2 36 B26 WX036 ##STR00136##
.sup.1H NMR (400 MHz, DMSO-d.sub.6)= 11.00 (s, 2H), 9.22 (s, 2H),
8.99 (s, 1H), 8.69 (d, J = 2.8 Hz 1H), 8.17 (s, 1H), 7.88 (s, 1H),
7.70 (s, 1H), 7.65 (d, J = 5.2 Hz 1H), 7.45-7.41 (m, 3H), 6.99 (d,
J = 4.4 Hz 1H), 6.90- 6.85 (m, 1H), 2.77-2.73 (m, 2H), 2.61 (s,
3H), 1.82-1.77 (m, 2H), 1.16 (s, 6H). 516.2
Embodiment 37: Synthesis of Compound WX037
##STR00137##
[0162] Synthetic Route:
##STR00138##
[0163] Step 1: Synthesis of Compound WX037-1
[0164] 2-Amino-4-chloro-5-nitropyridine (10.0 g, 57.62 mmol) and
ethyl bromoacetyl pyruvate (21.5 g, 97.95 mmol) were added to a
reaction flask, and the mixture was replaced with nitrogen three
times, and the reaction was stirred at 110.degree. C. for 16 hours.
Ethanol (80 mL) was added to the reaction mixture, and the mixture
was stirred for 2 hours, then filtered; the solid was collected,
and concentrated under reduced pressure to dryness to obtain
compound WX037-1.
[0165] Step 2: Synthesis of Compound WX037-2
[0166] WX037-1 (10.0 g, 37.1 mmol) and anhydrous ethanol (100.0 mL)
were added to a reaction flask, then concentrated sulfuric acid
(3.7 g, 37.1 mmol, 2.0 mL, 98% purity) was added thereto, and the
reaction mixture was stirred at 80.degree. C. for 12 hours. The
reaction mixture was concentrated under reduced pressure. Ethyl
acetate (300.0 mL) was added for dissolution, then pH value was
adjusted to 8 with saturated sodium carbonate aqueous solution; the
phases were separated, and the organic phase was dried over
anhydrous sodium sulfate, filtered and concentrated under reduced
pressure. The crude product was purified by column chromatography
(dichloromethane:methanol=100: 0-10:1) to obtain WX037-2.
[0167] Step 3: Synthesis of Compound WX037-3
[0168] WX037-2 (7.5 g, 25.2 mmol) and isopropyl acetate (140.0 mL)
were added to a reaction flask, then stannic chloride dihydrate
(34.1 g, 151.2 mmol) was added thereto. The mixture was stirred at
50.degree. C. for 12 hours. Ethyl acetate (200.0 mL) was added to
the reaction mixture, then the pH value was adjusted to 9 by adding
ammonia water dropwise, and anhydrous sodium sulfate was added, and
the sodium sulfate was stirred into a sand form; the mixture was
filtered, and the filtrate was collected and concentrated to
dryness under reduced pressure. The crude product was purified by
column chromatography (eluent: dichloromethane:methanol=100:
0-10:1) to obtain WX037-3.
[0169] Step 4: Synthesis of Compound WX037-4
[0170] A mixture of WX037-3 (3 g, 11.21 mmol, 1 eq), A2 (3.0 g,
14.6 mmol), N,N-diisopropylethylamine (5.8 g, 44.8 mmol, 7.8 mL),
50% ethyl acetate solution of tri-n-propyl cyclophosphoric
anhydride (21.9 g, 33.6 mmol, 20.0 mL, 50% purity) and THF (50.0
mL) was added to a reaction flask. The mixture was stirred at
50.degree. C. for 12 hours. Ethyl acetate (100.0 mL) was added,
then the pH value was adjusted to 8 with saturated sodium carbonate
aqueous solution; the phases were separated, and the organic phase
was collected and dried over anhydrous sodium sulfate, filtered and
concentrated under reduced pressure. The crude product was purified
by column chromatography (dichloromethane:methanol=100: 0-10:1) to
obtain WX037-4.
[0171] Step 5: Synthesis of Compound WX037-5
[0172] WX037-4 (1.0 g, 2.2 mmol), B19 (486.8 mg, 2.9 mmol),
potassium phosphate (1.4 g, 6.6 mmol) [methanesulfonic acid
(2-dicyclohexylphosphine)-3,6-dimethoxy-2,4,6-triisopropyl-1,1-biphenyl)(-
2-amino-1,1-biphenyl-2-yl) palladium (II) (299.6 mg, 330.5
.mu.mol), tetrahydrofuran (10.0 mL) and water (3.0 mL) were added
to a reaction flask, then the mixture was replaced with nitrogen
three times, and stirred at 80.degree. C. for 12 hours. Then the
mixture was filtered, and the filtrate was collected, then
concentrated to dryness under reduced pressure. The crude product
was purified by column chromatography
(dichloromethane:methanol=100: 0-10:1) to obtain WX037-5.
[0173] Step 6: Synthesis of Compound WX037
[0174] WX037-5 (0.05 g, 92.9 .mu.mol), sodium hydroxide (2 M, 919.9
.mu.L), methanol (5.0 mL) were added to a reaction flask, and the
mixture was replaced with nitrogen three times, then stirred at
25.degree. C. for 2 hours. Methanol was concentrated to dryness
under reduced pressure, then the pH was adjusted to 7 with 2N
hydrochloric acid, and then the mixture was concentrated to dryness
under reduced pressure. Then the crude product was purified by
machine purification (column: Phenomenex Gemini NX-C18 (75*30 mm*3
.mu.m); mobile phase: [aqueous solution containing (10.0 mM)
NH.sub.4HCO.sub.3)-acetonitrile]; gradient: B %: 15%-40%, 8 min) to
obtain WX037.
[0175] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.=11.5 (s, 1H),
9.21 (s, 1H), 9.02 (s, 1H), 8.98 (s, 1H), 8.70-8.69 (d, J=4.0 Hz,
1H), 7.90 (s, 1H), 7.65-7.58 (m, 3H), 7.49 (s, 1H), 7.17-7.15 (d,
J=8.0 Hz, 1H), 7.09-7.04 (t, 1H), 3.80 (s, 3H), 2.97-2.93 (t, 2H),
2.69-2.65 (t, 2H), 2.61 (s, 3H).
[0176] LCMS (ESI) m/z: 516.1[M+H].sup.+.
[0177] Each of the embodiments in the following Table 6 was
synthesized with reference to the synthesis step of embodiment 37,
except that the B-19 in step 5 was replaced with Fragment 1.
TABLE-US-00006 TABLE 6 Embodi- Frag- MS m/z ments ment 1 Compound
Product structure NMR [M + H].sup.+ 38 B16 WX038 ##STR00139##
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. = 12.19-11.96 (m, 1H),
9.48 (s, 1H) 9.02 (s, 1H), 8.91 (s, 1H), 8.67 (s, 1H), 7.88 (s,
1H), 7.69- 7.62 (m, 1H), 7.55-7.51 (m, 1H), 7.39-7.37 (m, 2H), 7.35
(s, 2H), 2.99- 2.95 (m, 2H), 2.70-2.66 (m, 2H), 2.60 (s, 3H) 2.33
(s, 1H). 486.3 39 B10 WX039 ##STR00140## .sup.1H NMR (400 MHz,
DMSO-d.sub.6)= 11.5 (s, 1H), 9.71 (s, 1H), 9.00 (s, 1H), 8.92 (s,
1H), 8.68-8.67 (d, J = 4.0 Hz, 1H), 7.82 (s, 1H), 7.72 (s, 1H),
7.66-7.64 (m, 1H), 7.56-7.46 (m, 6H), 2.96-2.93 (m, 2H), 2.68- 2.64
(m, 2H), 2.59 (s, 3H). 468.1
Embodiment 40: Synthesis of Compound WX040
##STR00141##
[0179] Synthetic Route:
##STR00142##
[0180] Step 1: Synthesis of Compound WX040-1
[0181] WX001-1 (5.4 g, 24.3 mmol) was added to a flask containing
ethyl bromopyruvate (47.4 g, 243.0 mmol, 30.4 mL) and the reaction
mixture was stirred at 90.degree. C. for 12 hours. The reaction
mixture was poured into ethyl acetate (150.0 mL) while hot, then
stirred at 15.degree. C. for 15 minutes, filtered under suction,
and the filter cake was rinsed with ethyl acetate (20.0
mL.times.3), and concentrated to dryness under reduced pressure to
obtain WX040-1.
[0182] Step 2: Synthesis of Compound WX040-2
[0183] Raney nickel (942.0 mg) was added to an argon-protected
hydrogenation flask, then the flask was wetted with ethanol (30.0
mL), and WX040-1 (1.0 g, 3.1 mmol) was added to the reaction
system; the mixture was stirred at 25.degree. C. for 2 hours under
50 Psi hydrogen. The reaction mixture was filtered through celite
under suction, and the filtrate was concentrated to dryness under
reduced pressure to obtain WX040-2.
[0184] Step 3: Synthesis of Compound WX040-3
[0185] WX040-2 (200 mg, 693.6 .mu.mol), A2 (170.0 mg, 832.34
.mu.mol), O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium
hexafluorophosphate (395.6 mg, 1.0 mmol), N,N-diisopropylethylamine
(268.9 mg, 2.1 mmol, 362.4 .mu.L) were added to a flask containing
anhydrous dichloromethane (15.0 mL), and the reaction mixture was
stirred at 25.degree. C. for 2 hours. The reaction mixture was
poured into saturated ammonium chloride solution (20.0 mL); the
phases were separated, and the organic phase was dried, filtered
and concentrated under reduced pressure. The crude product was
eluted by column separation (petroleum ether to petroleum
ether:ethyl acetate=1:1 to pure ethyl acetate) to obtain
WX040-3.
[0186] Step 4: Synthesis of Compound WX040
[0187] Methyl magnesium chloride (3.0 mol/L, 4.2 mL) was added to a
flask containing anhydrous tetrahydrofuran (15.0 mL) under nitrogen
protection, and WX040-3 (100 mg, 210.7 .mu.mol) dissolved in
anhydrous tetrahydrofuran (9.0 mL) was added dropwise to the above
solution at 20.degree. C., and the reaction was stirred at
20.degree. C. for 0.5 hours. The reaction mixture was quenched by
pouring into saturated ammonium chloride solution (20.0 mL), and
the mixture was extracted with ethyl acetate (10.0 mL.times.4); the
organic phases were dried, filtered and concentrated under reduced
pressure. The crude product was purified by plate separation (ethyl
acetate:methanol=10:1) to obtain WX040.
[0188] .sup.1H NMR (400 MHz, CD.sub.3OD-d.sub.4) .delta.=9.41 (s,
1H), 8.75 (s, 1H), 8.66 (d, J=5.2 Hz 1H), 7.99 (s, 1H), 7.90 (d,
J=5.6 Hz 1H), 7.66 (s, 1H), 7.20 (s, 1H), 3.15-2.99 (m, 4H), 2.67
(s, 3H), 3.11-1.98 (m, 4H), 1.85-1.64 (m, 2H), 1.62 (s, 6H).
[0189] LCMS (ESI) m/z: 461.3 [M+H].sup.+.
[0190] Each of the embodiments in the following Table 7 was
synthesized with reference to the synthesis step of embodiment 21
and embodiment 40, except that the piperidine at the bottom right
was replaced with Fragment 1.
TABLE-US-00007 TABLE 7 Embodi- Frag- MS m/z ments ment 1 Compound
Product structure NMR [M + H].sup.+ 41 B25 WX041 ##STR00143##
.sup.1H NMR (400 MHz, DMSO-d.sub.6)= 10.03 (s, 1H), 8.94 (s, 1H),
8.70 (s, 1H), 8.69 (s, 1H), 7.86 (s, 1H), 7.81 (m, 1H), 7.73- 7.72
(s, 1H), 7.62 (s, 1H), 7.34-7.30 (m, 3H), 5.15 (s, 1H), 2.60 (s,
3H), 1.51 (s, 6H). 490.1
Embodiment 42: Synthesis of Compound WX042
##STR00144##
[0192] Synthetic Route:
##STR00145##
[0193] WX042-1 was synthesized with reference to the synthesis step
of embodiment 1, except that the A1 in step 2 was replaced with
ethyl 4-bromoacetoacetate.
[0194] Step 1: Synthesis of Compound WX042-2
[0195] Compound WX042-1 (0.8 g, 1.64 mmol) was dissolved in
tetrahydrofuran (10.0 mL), and the mixture was cooled to
-10.degree. C.; lithium aluminum tetrahydride (155.4 mg) was added
in batches to the reaction system, and the reaction was stirred at
-10.degree. C. for 1 hour. The reaction mixture was poured into
ammonium chloride aqueous solution (50.0 mL), and the mixture was
extracted with ethyl acetate (50.0 mL.times.4), and the organic
phases were combined, washed with saturated saline (100.0 mL),
dried over anhydrous sodium sulfate, filtered and concentrated
under reduced pressure. The crude product was purified by column
purification (dichloromethane:methanol=100: 0-100:0.25) to obtain
WX042-2.
[0196] Step 2: Synthesis of Compound WX042-3
[0197] Compound WX042-2 (200.0 mg) was dissolved in chloroform
(10.0 mL), and triethylamine (136.0 mg) was added thereto, then the
mixture was cooled to 10.degree. C. and stirred for 10 minutes,
then a solution of methanesulfonyl chloride (77.0 mg) in chloroform
(1.0 mL) was slowly added dropwise. The reaction was slowly heated
to 25.degree. C. and stirred for 20 minutes. The reaction mixture
was concentrated under reduced pressure to obtain WX042-3.
[0198] Step 3: Synthesis of Compound WX042
[0199] Compound WX042-3 (0.2 g) and sodium methylsulfinate (70.1
mg, 686.3 .mu.mol) were dissolved in N,N-dimethylformamide (10.0
mL), and potassium iodide (189.8 mg) was added thereto. The
reaction was carried out at 80.degree. C. (0 bar) for 1 hour in
microwave instrument. The reaction mixture was diluted with 10.0 mL
of ethyl acetate, then the mixture was poured into semi-saturated
saline (50.0 mL); the phases were separated, and the aqueous phase
was extracted with ethyl acetate (50.0 mL.times.4), and the organic
phases were combined, dried over anhydrous sodium sulfate, filtered
and concentrated under reduced pressure. Then the crude product was
purified by machine purification (column: Welch Xtimate BEH C18
100*30 mm*10 .mu.m; phase: A: aqueous solution containing 10 mM
NH.sub.4HCO.sub.3, B: acetonitrile; gradient: B %: 30%-50%, 6
minutes) and freeze-dried to obtain WX042.
[0200] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.=9.87 (s, 1H),
9.35 (s, 1H), 9.09 (s, 1H), 8.71 (s, 1H), 7.83 (s, 2H), 7.73 (s,
1H), 7.23 (s, 1H), 3.48-3.46 (m, 2H), 3.44 (s, 2H), 3.09-2.99 (m,
3H), 2.91-2.90 (m, 4H), 2.60-2.58 (m, 3H), 1.87-1.86 (m, 4H), 1.67
(s, 2H)
[0201] LCMS (ESI) m/z: 509.1 [M+H].sup.+.
[0202] Each of the embodiments in the following Table 8 was
synthesized with reference to the synthesis step of embodiment 42,
except that the piperidine at the bottom right was replaced with
Fragment 1.
TABLE-US-00008 TABLE 8 Embodi- Frag- MS m/z ments ment 1 Compound
Product structure NMR [M + H].sup.+ 43 B25 WX043 ##STR00146##
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. = 9.96 (s, 1H), 8.91
(s, 2H), 8.67 (d, J = 5.2 Hz 1H), 7.93 (s, 1H), 7.78 (s, 1H), 7.69
(d, J = 4.0 Hz 1H), 7.61 (s, 1H), 7.33-7.29 (m, 3H), 3.55-3.48 (m,
2H), 3.20- 3.15 (m, 2H), 3.01 (s, 3H), 2.61 (s, 3H). 538.1
Test Embodiment 1: Evaluation of IRAK4 Kinase Activity In Vitro
[0203] The IC.sub.50 values were determined using .sup.33P
isotope-labeled kinase activity assay (Reaction Biology Corp) to
evaluate the inhibitory ability of the tested compounds on human
IRAK4.
[0204] Buffer conditions: 20 mM Hepes (pH 7.5), 10 mM MgCl.sub.2, 1
mM EGTA, 0.02% Brij35, 0.02 mg/mL BSA, 0.1 mM Na.sub.3VO.sub.4, 2
mM DTT, 1% DMSO.
[0205] Test procedure: At room temperature, the tested compound was
dissolved in DMSO to prepare a 10 mM solution for later use. The
substrate was dissolved in a newly prepared buffer solution, and
the tested IRAK4 kinase was added thereto and mixed evenly. The
DMSO solution dissolved with the tested compound was added to the
above reaction mixture mixed evenly using acoustic technique (Echo
550). After incubation for 15 minutes, .sup.33P-ATP was added to
initiate the reaction. The reaction was carried out at room
temperature for 120 minutes, and the reaction mixture was spotted
on P81 ion exchange filter paper (Whatman #3698-915). The filter
paper was washed repeatedly with 0.75% phosphoric acid solution,
and the radioactivity of phosphorylated substrate residues on the
filter paper was determined. The kinase activity data were
expressed by comparing the kinase activity of the group containing
the tested compound with the kinase activity of the blank group
(only containing DMSO), and the IC.sub.50 value was obtained by
curve fitting by Prism4 software (GraphPad), and the experimental
results are shown in Table 9.
TABLE-US-00009 TABLE 9 In vitro IRAK4 kinase activity screening
test results of the compounds of the present disclosure Number of
the compound IRAK4/IC.sub.50 (nM) WX001 1.0 WX002 1.2 WX003 2.2
WX004 1.3 WX005 1.1 WX006 1.2 WX007 2.0 WX008 2.1 WX009 0.7 WX010
2.4 WX011 1.5 WX012 1.0 WX013 0.2 WX014 0.9 WX015 0.4 WX016 0.9
WX017 1.6 WX018 0.9 WX020 0.5 WX021 0.5 WX022 0.7 WX023 8.4 WX024
0.7 WX025 0.7 WX027 11 WX033 1.6 WX039 1 WX040 1.2 WX044 1.9 WX045
3.4 WX046 2.5
[0206] Conclusion: The compound of the present disclosure generally
exhibits good inhibitory activity against IRAK4.
Test Embodiment 2: Evaluation of BTK Kinase Activity In Vitro
[0207] The IC.sub.50 values were determined using .sup.33P
isotope-labeled kinase activity assay (Reaction Biology Corp) to
evaluate the inhibitory ability of the tested compounds on human
BTK.
[0208] Buffer conditions: 20 mM Hepes (pH 7.5), 10 mM MgCl.sub.2, 1
mM EGTA, 0.02% Brij35, 0.02 mg/mL BSA, 0.1 mM Na.sub.3VO.sub.4, 2
mM DTT, 1% DMSO.
[0209] Test procedure: At room temperature, the tested compound was
dissolved in DMSO to prepare a 10 mM solution for later use. The
substrate was dissolved in a newly prepared buffer solution, and
the tested BTK kinase was added thereto and mixed evenly. The
compound dissolved in DMSO was added to the kinase reaction mixture
through Echo 550 (Acoustic technology; Nanoliter range). After
incubation for 20 minutes at room temperature, .sup.33P-ATP was
added to initiate the reaction. The reaction was carried out at
room temperature for 2 hours, and the radioactivity of the reaction
liquid point was detected by filtration-binding method with P81 ion
exchange filter paper. The kinase activity data were expressed by
comparing the kinase activity of the group containing the tested
compound with the kinase activity of the blank group (only
containing DMSO), and the IC.sub.50 value was obtained by curve
fitting by Prism4 software (GraphPad), the experimental results are
shown in Table 10.
TABLE-US-00010 TABLE 10 In vitro BTK kinase activity screening test
results of the compoundsof the present disclosure Number of the
compound BTK/IC.sub.50 (nM) WX001 6.7 WX006 47.5 WX008 4.6 WX009 46
WX011 23.6 WX012 6.9 WX013 1.4 WX014 25.7 WX015 6.4 WX016 8.9 WX018
47.5 WX020 8.5 WX021 13.9 WX022 43.1 WX024 30.5 WX025 21.8 WX026
30.7 WX029 24.2 WX031 39.6 WX032 26.5 WX033 20.2 WX039 29 WX040 3.3
WX042 3.8 WX045 28.6 WX046 43.1
[0210] Conclusion: The compound of the present disclosure generally
exhibits good inhibitory activity against BTK.
Test Embodiment 3: Evaluation of THP-1 Cytological Activity In
Vitro
[0211] THP-1 Cytological TNFa ELISA Assay
[0212] 1. Experimental Materials:
[0213] THP-1 human acute single cell leukemia cells were purchased
from ATCC (Cat #TIB-202) and cultured at 37.degree. C. in 5%
CO.sub.2 incubator. The composition of medium was RPMI1640 (Gibco,
Cat #22400-105), and the supplementary compositions were 10% FBS
(Gibco, Cat #10091148); 1% PenStrep (Gibco, Cat #15140); 0.05 mM
2-Mercaptoethanol (Sigma, Cat #M6250).
[0214] 2. Experimental Methods:
[0215] TNF-.alpha. Elisa kit was used to detect the content of
TNF-.alpha. in cell culture supernatant samples. TNF-.alpha. was
produced by stimulating THP-1 cells with 150 ng/mL LPS (Sigma, Cat
#L6529).
[0216] THP-1 cells cultured normally at logarithmic growth stage
were seeded in a 96-well plate (Corning #3599) at a certain
concentration (1*10.sup.5/100 .mu.L) and then put into a cell
incubator for incubation. After two hours, 16.7 .mu.L of different
concentrations of the compound to be tested (8*final concentration)
were added and incubated in an incubator. After one hour, 16.7
.mu.L of 1200 ng/mL LPS was added and incubated in an incubator.
After 18 hours, the culture supernatant samples were collected by
centrifugation, and the content of TNF-.alpha. could be detected by
TNF-.alpha. Elisa kit. Finally, OD signals (OD450-OD570) were read
on envision board reader.
[0217] 3. Data Analysis:
[0218] The OD450-OD570 signal value was converted into a percentage
inhibition rate.
Inhibition rate %=(ZPE-sample)/(ZPE-HPE)*100.
[0219] "HPE" refers to the OD450-OD570 signal value of the control
well without LPS stimulated cells, and "ZPE" refers to the
OD450-OD570 signal value of the control well with LPS stimulated
cells. The IC.sub.50 value of the compound was calculated by XLFit
in the excel add-in.
Y=Bottom+(Top-Bottom)/(1+(IC.sub.50/X){circumflex over (
)}HillSlope). Equation:
[0220] A summary of the test results is shown in Table 11.
TABLE-US-00011 TABLE 11 In vitro screening test results of the
compounds of the present disclosure Number of the compound
THP-1/IC.sub.50 (nM) WX001 124 WX003 557 WX004 77 WX005 256 WX006
107 WX007 240 WX010 332 WX011 140 WX016 170 WX017 321 WX018 104
WX021 15 WX024 13 WX025 21 WX040 38
[0221] Conclusion: The compound of the present disclosure generally
exhibits better activity of inhibiting cell TNF-.alpha. generation
in THP-1 cell activity experiment.
Test Embodiment 4: Evaluation of OCI-LY10 and TMD-8 Cytological
Activity In Vitro
[0222] 1. Experimental Materials:
[0223] OCI-LY10 human B-cell lymphoma cells were cultured in a
37.degree. C., 5% CO.sub.2 incubator. The composition of medium was
IMDM (GIBCO, Cat #12440053); the supplementary compositions were
20% FBS (Hyclone, Cat #SH30084.03); 1% PenStrep (Thermo, Cat
#SV30010).
[0224] TMD8 human B-cell lymphoma cells were cultured in a
37.degree. C., 5% CO.sub.2 incubator. The composition of medium was
RPMI1640 (GIBCO, Cat #22400-089); the supplementary compositions
were 10% FBS (Hyclone, Cat #SH30084.03); 1% PenStrep (Thermo, Cat
#SV30010).
[0225] 2. Experimental Methods:
[0226] The tumor cell lines OCI-LY10 and TMD8 were used to detect
the effect of the compound on inhibiting tumor cell proliferation
in vitro. The tumor cell line was cultured in a 37.degree. C., 5%
CO.sub.2 incubator according to the culture conditions shown, and
passaged regularly, then the cells in the logarithmic growth phase
were taken, counted, and spread in a 96-well plate (the cells in
each well were adjusted to an appropriate concentration, a total of
90 cell suspensions per well was added). After incubating overnight
in a 37.degree. C., 5% CO.sub.2 incubator, drugs with different
concentration gradients (10 .mu.L of drug solution was added) were
added and treated for 3 days, then 50 .mu.L of CellTiter-Glo
working solution was added to each well, and the cell plate was
wrapped with aluminum foil to avoid light. The culture plate was
shaken on an orbital shaker for 2 minutes to induce cell lysis, and
placed at room temperature for 10 minutes to stabilize the
luminescence signal, then the luminescence signal was detected on
the 2104 EnVision plate reader.
[0227] 3. Data Analysis:
[0228] The Inhibition rate (IR) of the tested compound was
calculated using the following formula:
IR (%)=(1-(RLU compound-RLU blank control)/(RLU solvent control-RLU
blank control))*100%.
[0229] The inhibition rates of different concentrations of the
compounds were calculated in Excel, and then the inhibition curves
were made by GraphPad Prism software and the related parameters
were calculated, including the minimum inhibition rate, the maximum
inhibition rate and IC.sub.50.
[0230] 4. Experimental Results
[0231] Experimental results are shown in Table 12:
TABLE-US-00012 TABLE 12 In vitro screening test results of the
compounds of the present disclosure Number of the compound
OCI-LY10/IC.sub.50 (nM) TMD-8/IC.sub.50 (nM) WX001 128 300 WX016
466 221 WX021 171 215 WX024 208 / WX025 94 / WX026 349 / WX029 289
/ WX030 209 / WX031 96 / WX032 386 / WX040 87 / WX045 77 /
[0232] Conclusion: The compounds of the present disclosure
generally exhibit good inhibitory activity on cell proliferation in
OCI-LY10 and TMD-8 cell lines, respectively.
[0233] Note: "/" means not detected.
Test Embodiment 5: Evaluation of OCI-LY3 Cytological Activity In
Vitro
[0234] 1. Experimental Cell Line Information and Cell Culture
[0235] The tumor cell line used in this experiment was provided by
Nanjing Cobioer Biotechnology Co., Ltd. See Table 13 below for
specific information.
TABLE-US-00013 TABLE 13 Experimental cell line information Cell
name Cell source Cell culture medium OCI-LY3 Nanjing Cobioer
Biotechnology Co., IMDM+20% FBS+0.05 l Ltd mM 2-mercaptoethano +1%
penicillin/streptomycin
[0236] 2. Experimental Methods:
[0237] The tumor cell lines OCI-LY3 were used to detect the effect
of the compound on inhibiting tumor cell proliferation in vitro.
The OCI-LY3 cell line was cultured in the corresponding medium at
37.degree. C. and 5% CO.sub.2, and the logarithmic growth phase
cells were used in the experimental plating. The cells were
collected and centrifuged at 800 rpm for 5 minutes, and the culture
medium was re-suspended and spread in a 96-well plate. After
incubating overnight in a 37.degree. C., 5% CO.sub.2 incubator, the
cells with different concentration gradients (10 .mu.L of prepared
diluent of the tested compound) were incubated for 72 hours, and
the cell culture plates were incubated with CTG reagent at room
temperature and away from light for 30 minutes, and then recovered
to room temperature. 100 .mu.L/hole of CTG solution was added into
the biosafety cabinet away from light, and the plate shaker was
shaken and mixed evenly away from light for 2 minutes, and
incubated at room temperature away from light for 10 minutes. The
luminescence values were read and recorded using the Perkin Elmer
Envision 2104 MuLtilabel Reader.
[0238] 3. Data Processing and Analysis
[0239] The results of luminescence values measured at each drug
concentration were normalized with the luminescence values of the
blank control group, and the ratio of this value to the DMSO group
was taken as the cell inhibition rate (%). Using GraphPad software,
the logarithm of drug concentration (log drug concentration) versus
inhibition rate was plotted, and the software automatically fitted
and calculated IC.sub.50 value and 95% confidence limit value by
log (inhibitor) vs. normalized response algorithm of nonlinear
regression.
[0240] 4. Experimental Results
[0241] Experimental results are shown in Table 14.
TABLE-US-00014 TABLE 14 In vitro screening test results of the
compounds of the present disclosure Number of the compound OCI-LY3/
IC.sub.50 (uM) WX001 1.292 WX040 0.283 WX045 0.270
[0242] Conclusion: The compound of the disclosure has a significant
inhibition effect on cell proliferation in OCI-LY3 cell line.
Test Embodiment 6: In Vivo Pharmacodynamic Study of TNF-.alpha.
Secretion in SD Rats Induced by Lipopolycollagen (LPS)
[0243] 1. Modeling and Administration
[0244] SD rats were orally given the solvent, the positive drug
dexamethasone (DEX, 0.5 mg/kg) and the tested compound, and LPS (1
mg/kg) was intraperitoneally injected 0.5 hours after the
administration. Two hours after LPS injection, the animals were
euthanized by CO.sub.2, and blood samples were collected from the
heart and placed in an anticoagulant tube containing EDTA-K.sub.2,
then partial anticoagulated blood was centrifuged to separate the
plasma and the plasma was frozen at -80.degree. C.
[0245] 2. TNF-.alpha. Detection
[0246] The plasma was taken out of the refrigerator at -80.degree.
C., thawed at room temperature, and the concentration of
TNF-.alpha. in the plasma was detected according to the ELISA kit
instructions.
[0247] 3. Statistical Processing
[0248] The experimental data were expressed by Mean.+-.standard
error (Mean.+-.SEM), and the level of TNF-.alpha. was expressed by
One-way ANOVA, and p<0.05 was considered as a significant
difference. The result of in vivo pharmacodynamic study of
TNF-.alpha. secretion in SD rats induced by lipopolycollagen (LPS)
are shown in FIG. 1
[0249] 4. Experimental Results
[0250] The results in FIG. 1 show that the SD rat orally
administrated compound WX001 showed a significant inhibitory effect
on TNF-.alpha. secretion induced by lipopolycollagen (LPS), and the
efficacy at a dose of 20 mpk was equivalent to the efficacy of
dexamethasone (DEX) at a dose of 0.5 mpk.
Test Embodiment 7: In Vivo Pharmacodynamic Study of WX001 on Human
B-Cell Lymphoma OCI-LY10 Cell Subcutaneous Xenograft Tumor Mouse
Model
[0251] 1. Experimental Objectives
[0252] The objective of the experiment was to study the efficacy of
WX001 as the test drug on human B-cell lymphoma OCI-LY10 cell
subcutaneous xenograft tumor in CB17 SCID mouse model.
[0253] 2. Experimental Materials
[0254] OCI-LY10 human B-cell lymphoma cells were cultured in a
37.degree. C., 5% CO.sub.2 incubator. The composition of medium was
IMDM (GIBCO, Cat #12440053); the supplementary compositions were
20% FBS (Hyclone, Cat #SH30084.03); 1% PenStrep (Thermo, Cat
#SV30010).
[0255] 3. Experimental Methods
[0256] OCI-LY10 tumor cells were cultured and passaged, and 0.2 mL
(1.times.10.sup.7 cells) OCI-LY10 cells were subcutaneously
inoculated on the right back of each nude mouse (with Matrigel,
volume ratio 1:1), and the group administration was started when
the average tumor volume reached 167 mm.sup.3. The health status
and death of animals were monitored every day, and routine
examinations included observing the effects of tumor growth and
drug treatment on daily behaviors of animals, such as behavioral
activities, food intake and water intake, weight change (weight was
measured twice a week), tumor size (tumor volume was measured twice
a week), appearance signs or other abnormal conditions.
[0257] 4. Data Analysis
[0258] The experimental index was to investigate whether the tumor
growth was inhibited, delayed or cured. Including the measurement
of tumor volume (TV), and the calculation of the compound's
anti-tumor efficacy using TGI (%) or the relative tumor
proliferation rate T/C (%).
[0259] TV=0.5a.times.b.sup.2, a and b represented the long diameter
and short diameter of the tumor, respectively.
[0260] TGI (%)=(1-(average tumor volume at the end of
administration in a treatment group-average tumor volume at the
beginning of administration in the treatment group))/(average tumor
volume at the end of treatment in solvent control group-average
tumor volume at the beginning of treatment in solvent control
group)).times.100%.
[0261] T/C %=T.sub.RTV/C.sub.RTV.times.100% (T.sub.RTV: RTV in
treatment group; C.sub.RTV: RTV in negative control group).
Relative tumor volume (RTV) was calculated according to the results
of tumor measurement, and the calculation formula was
RTV=V.sub.t/V.sub.0, wherein V.sub.0 was the average tumor volume
measured at the time of group administration (i.e., d.sub.0),
V.sub.t was the average tumor volume at a certain measurement, and
T.sub.RTV and C.sub.RTV were the data taken at the same day.
[0262] 5. Experimental Results
[0263] 5.1. Mortality, Morbidity and Weight Changes
[0264] The body weight of experimental animals was used as a
reference index for indirect determination of drug toxicity. After
18 days of administration (PG-D1-D18), all mice in the experimental
group showed no abnormality and showed good drug tolerance.
[0265] The effect of WX001 compound on the body weight of female
CB17 SCID mouse model bearing human B-cell lymphoma OCI-LY10 cell
subcutaneous xenograft tumor is shown in FIG. 2 and FIG. 3. FIG. 2
shows the weight changes of mouse model bearing human B-cell
lymphoma OCI-LY10 cell subcutaneous xenograft tumor after
administration of WX compound. The data points represent the
average body weight in the group, and the error lines represent the
standard error (SEM). The relative weight change shown in FIG. 3
was calculated based on the animal weight at the beginning of
administration. The data points represent the average body weight
change percentage in the group, and the error lines represent the
standard error (SEM).
[0266] 5.2. Tumor Growth Curve
[0267] FIG. 4 shows the tumor growth curve of mouse model bearing
human B-cell lymphoma OCI-LY10 cell subcutaneous xenograft tumor
after administration of WX001 compound. The data points represent
the average tumor volume in the group, and the error lines
represent the standard error (SEM).
[0268] 6. Experimental Results and Discussion
[0269] In this study, we evaluated the in vivo efficacy of WX001
compound in human B-cell lymphoma OCI-LY10 cell subcutaneous
xenograft tumor model. The tumor volume of each group at different
time points is shown in FIG. 4.
[0270] 18 days after the start of administration, the T/C value of
the Ibrutinib (10 mpk) group was 39%, and the TGI value was 85%,
and the p value was <0.001. The WX001 (50 mpk) group had a T/C
value of 22%, a TGI value of 109%, and p<0.001; compared with
the solvent control group, the WX001 (50 mpk) group had a
significant anti-tumor effect and was significantly better than the
Ibrutinib (10 mpk) group.
[0271] The OCI-LY10 cell line is an ABC-DLBCL cell line that is
highly dependent on both MyD88-L265P and BCR (CD79A/B) double
mutations. IRAK4 and BTK dual-target inhibitor WX001 (50 mpk) shows
the significant anti-tumor effect as a single agent (TGI=109%),
which is significantly better than the single-agent efficacy of
Ibrutinib (10 mpk) (TGI=85%), shows significant simultaneous
IRAK4/BTK pathway inhibition, and it is well tolerated by
animals.
* * * * *